Cell Biology | DNA Transcription 🧬

01:25:28
https://www.youtube.com/watch?v=ZrCx98CtJ_4

الملخص

TLDRVideon från Ninja Nerd handlar om DNA-transkription, vilket är processen där DNA omvandlas till RNA. Den täcker reglerande element och enzymer som deltar i processen, såsom RNA-polymeras och transkriptionsfaktorer. För prokaryoter sker transkriptionen med ett RNA-polymeras som består av core enzym och en sigma-enhet medan eukaryoter använder tre olika RNA-polymeraser och flera allmänna transkriptionsfaktorer. Videon diskuterar även post-transkriptionella modifieringar som capping, poly-A svans och splitsning av introner och exoner. Alternativ RNA-splitsning förklaras, där samma gen kan leda till olika proteiner beroende på hur mRNA splitsas. Skillnader i transkriptionella processer mellan prokaryoter och eukaryoter tas också upp, inklusive rho-beroende och rho-oberoende terminering samt eukaryotisk genreglering med enhancers och silencers. Videon avslutas med en diskussion om RNA-redigering, där exempel ges på hur samma RNA-sekvens kan modifieras för att producera olika proteiner i olika celltyper.

الوجبات الجاهزة

  • 📘 Förståelse av DNA-transkription och dess definition.
  • 🔍 Skillnad mellan prokaryotisk och eukaryotisk transkription.
  • 🧬 Roller hos RNA-polymeras i transkription.
  • 📍 Vikt av promotorregioner i transkription.
  • 🛠 Funktion av 5'-cap och poly-A-svans i mRNA.
  • 🔄 Alternativ RNA-splitsning och dess betydelse.
  • 🚀 Roll av enhancers och silencers i genreglering.
  • 🛑 Mekanismer för terminering av transkription.
  • 🧪 Post-transkriptionella modifieringar i eukaryoter.
  • 🌱 RNA-redigering och dess biologiska konsekvenser.

الجدول الزمني

  • 00:00:00 - 00:05:00

    Introduktion till DNA-transkription. Vi kommer att förklara processen där DNA omvandlas till RNA i både eukaryota och prokaryota celler. För att denna process ska ske behövs vissa proteiner eller enzymer.

  • 00:05:00 - 00:10:00

    Fokus på prokaryota celler och deras behov av RNA-polymeras holoenzym, som består av kärnenzym och en sigma-subenhet, för transkriptionen. Promotorregionens roll i DNA beskrivs.

  • 00:10:00 - 00:15:00

    RNA-polymeras holoenzym läser DNA:s templatsträng för att syntetisera RNA. Prokaryota celler kan producera mRNA, rRNA och tRNA från en enda RNA-polymeras.

  • 00:15:00 - 00:20:00

    I eukaryota celler krävs specifika RNA-polymeraser och transkriptionsfaktorer. Tre RNA-polymeraser (I, II och III) ansvarar för produktionen av olika typer av RNA.

  • 00:20:00 - 00:25:00

    Transkriptionsfaktorer och deras roll i bindningen till promotorregioner i eukaryota celler beskrivs. RNA-polymeras I producerar rRNA, polymeras II producerar mRNA, och polymeras III producerar tRNA.

  • 00:25:00 - 00:30:00

    Processen för initiering av transkription i eukaryota celler: polymeras II och generella transkriptionsfaktorer binder till promotorregioner. Jämförelse mellan prokaryota och eukaryota system.

  • 00:30:00 - 00:35:00

    Elongering i transkription: RNA-polymeras öppnar och stabiliserar DNA-strängar och syntetiserar mRNA genom att läsa DNA-templatsträngen i 3'-5'-riktning.

  • 00:35:00 - 00:40:00

    Förklarar 5'-3'-läsning och syntes av RNA med hjälp av diagram för att illustrera processen.

  • 00:40:00 - 00:45:00

    Diskussion om hur RNA-polymerasers funktion kan hämmas av olika ämnen och vilka konsekvenser det får för cellerna. Termineringsprocessen i transkriptionen introduceras.

  • 00:45:00 - 00:50:00

    Termineringsprocessen i prokaryoter kan ske via rho-beroende eller rho-oberoende mekanismer. Beskrivning av hur rho-protein och hårnålsslingor påverkar processens avslutning.

  • 00:50:00 - 00:55:00

    Eukaryota celler använder polyadenyleringssignal för att avsluta transkriptionen. Enzymer klipper bort RNA från DNA och RNA-polymeraset.

  • 00:55:00 - 01:00:00

    Post-transkriptionella modifieringar i eukaryota celler innebär 5'-capping och poly-A-svans för stabilitet och transport av mRNA.

  • 01:00:00 - 01:05:00

    Cappning av RNA hjälper till att starta translation och skyddar mot nedbrytning. Poly-A-svansens roll diskuteras i liknande termer.

  • 01:05:00 - 01:10:00

    Splitsning av pre-mRNA i eukaryota celler tar bort introner och fogar samman exoner. Detta görs med hjälp av snurps, bestående av snRNA och proteiner.

  • 01:10:00 - 01:15:00

    Exon/intron strukturen i pre-mRNA beskrivs. Splitsningsprocessen avlägsnar introner och fogar samman exoner för att bilda moget mRNA.

  • 01:15:00 - 01:20:00

    Alternativ rna-splitsning ger möjlighet att producera olika proteiner från samma gen genom att variera exonkombinationer.

  • 01:20:00 - 01:25:28

    RNA-editering kan leda till olika proteinprodukter från samma mRNA, ett fenomen exemplifierat med apob100 och apob48 i enterocyter och hepatocyter.

اعرض المزيد

الخريطة الذهنية

Mind Map

الأسئلة الشائعة

  • Vad är DNA-transkription?

    DNA-transkription är processen där DNA omvandlas till RNA.

  • Vilka är de viktiga enzymerna i DNA-transkription för prokaryota celler?

    RNA-polymeras holoenzym med core enzym och sigma subenhet.

  • Vad är en promotorregion?

    En promotorregion är en nukleotidsekvens i DNA som tillåter bindning av RNA-polymeraser och transkriptionsfaktorer.

  • Vad är skillnaden mellan transkription i prokaryoter och eukaryoter?

    Prokaryoter använder ett RNA-polymeras, medan eukaryoter använder tre olika RNA-polymeraser och allmänna transkriptionsfaktorer.

  • Vad är alternatv RNA-splitsning?

    Alternativ RNA-splitsning är en process där samma gen kan ge upphov till olika proteinvarianter.

  • Varför är det viktigt med 5'-cap och poly-A-svans?

    De skyddar RNA från nedbrytning och hjälper till att initiera translation.

  • Vad är rho-beroende och rho-oberoende terminering?

    Rho-beroende terminering använder rho-proteinet för att avsluta transkriptionen, medan rho-oberoende bildar en hårnålsslinga som signalerar terminering.

  • Vad är post-transkriptionella modifieringar?

    Modifikationerna inkluderar 5'-capping, poly-A-svans läggs till och splitsning av exoner/introner.

  • Vad påverkar hastigheten på DNA-transkription i eukaryoter?

    Enhancers och silencers påverkar hastigheten på transkription genom att öka eller minska processen.

  • Vad är RNA-redigering?

    RNA-redigering är en process där nukleotider i RNA kan modifieras för att påverka proteintranslation.

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التمرير التلقائي:
  • 00:00:14
    what's up ninja nerds in this video
  • 00:00:15
    today we are going to be talking about
  • 00:00:17
    dna
  • 00:00:17
    transcription before we get started if
  • 00:00:19
    you guys do like this video please hit
  • 00:00:21
    that like button comment down in the
  • 00:00:22
    comment section
  • 00:00:23
    and please subscribe also down in the
  • 00:00:25
    description box we have links to our
  • 00:00:26
    facebook instagram patreon
  • 00:00:28
    all that stuff will be there all right
  • 00:00:29
    ninjas let's get into it all right ninja
  • 00:00:31
    so with dna transcription we have to
  • 00:00:33
    have a basic understanding of just the
  • 00:00:34
    definition what the heck is
  • 00:00:36
    transcription
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    and it's really a simple thing it's just
  • 00:00:38
    taking dna
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    okay double stranded dna with the
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    eukaryotic cells and even in prokaryotic
  • 00:00:43
    cells
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    and converting that into rna so it's
  • 00:00:46
    taking dna and making rna that's all
  • 00:00:48
    transcription is
  • 00:00:49
    but in order for transcription to occur
  • 00:00:52
    in order for it to take place
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    we need two particular types of proteins
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    or enzymes if you will
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    to facilitate this process and i want to
  • 00:01:00
    talk about those real quick
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    because these are very important now
  • 00:01:04
    transcription can be kind of different
  • 00:01:07
    okay and it's important to know the
  • 00:01:09
    differences
  • 00:01:09
    between prokaryotic cells we'll consider
  • 00:01:12
    bacteria in this case
  • 00:01:13
    and eukaryotic cells human cells like me
  • 00:01:15
    and you
  • 00:01:17
    in prokaryotic cells there's a
  • 00:01:19
    particular type of protein
  • 00:01:21
    that is needed in order for
  • 00:01:23
    transcription to take place
  • 00:01:25
    what is that protein so let's say that
  • 00:01:27
    we take
  • 00:01:28
    this dna strand here right we have this
  • 00:01:30
    dna strand
  • 00:01:31
    on this dna strand we have these blue
  • 00:01:33
    portions that i've highlighted here as a
  • 00:01:35
    box with some lines in it
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    this right here for right now i want you
  • 00:01:39
    to know is what's called a promoter so
  • 00:01:41
    this is called
  • 00:01:43
    a promoter region now a promoter region
  • 00:01:46
    is a
  • 00:01:47
    particular nucleotide sequence within
  • 00:01:49
    the dna
  • 00:01:51
    and what it does is it allows for
  • 00:01:53
    particular proteins like
  • 00:01:54
    rna polymerases and transcription
  • 00:01:56
    factors
  • 00:01:57
    to bind onto the dna and then start
  • 00:02:00
    moving through the dna
  • 00:02:01
    taking the dna and making rna so that's
  • 00:02:04
    the first thing you need to know is
  • 00:02:05
    within the dna
  • 00:02:07
    there's a particular nucleotide sequence
  • 00:02:08
    we'll talk about a little bit later
  • 00:02:10
    called a promoter region and that's the
  • 00:02:12
    first thing that we need to
  • 00:02:14
    identify let's say that we take this
  • 00:02:16
    particularly for
  • 00:02:19
    prokaryotic cells so prokaryotic cells
  • 00:02:23
    and we'll just say like a bacterial cell
  • 00:02:25
    okay prokaryotic cells use a
  • 00:02:28
    very particular type of enzyme what
  • 00:02:31
    is that enzyme it's called a
  • 00:02:34
    rna polymerase holoenzyme okay so it's
  • 00:02:38
    called an
  • 00:02:38
    rna we're going to put poll polymerase
  • 00:02:42
    holo enzyme
  • 00:02:46
    now that's a lot of stuff let me explain
  • 00:02:48
    what this is and i'll show you the
  • 00:02:49
    structure of it a basic structure of
  • 00:02:51
    what the rna polymerase holoenzyme is
  • 00:02:53
    it's made up of two things one of the
  • 00:02:56
    components of this enzyme
  • 00:02:59
    is called the core enzyme
  • 00:03:02
    and the core enzyme for this rna
  • 00:03:04
    polymerase holoenzyme
  • 00:03:07
    consists of multiple subunits that they
  • 00:03:09
    just love to ask you on your us mles and
  • 00:03:11
    other exams
  • 00:03:12
    and these are they contain two alpha
  • 00:03:15
    units
  • 00:03:16
    okay two alpha chains proteins it
  • 00:03:18
    contains
  • 00:03:19
    two beta units technically we say beta
  • 00:03:22
    and beta prime
  • 00:03:23
    if you really want to be specific and
  • 00:03:25
    then one more
  • 00:03:26
    which is called an omega unit okay
  • 00:03:29
    so these are the primary components of
  • 00:03:31
    the core enzyme which makes up rna
  • 00:03:33
    polymerase what's important to remember
  • 00:03:36
    is that these are what are going to
  • 00:03:38
    really read the dna
  • 00:03:39
    and make rna that portion of the enzyme
  • 00:03:42
    reads the dna
  • 00:03:43
    and makes rna the next component of the
  • 00:03:45
    rna polymerase holoenzyme is the portion
  • 00:03:48
    that we need in order to bind
  • 00:03:50
    to the dna to the promoter region
  • 00:03:51
    without it we won't be able to allow for
  • 00:03:53
    this rna polymerase to bind to the dna
  • 00:03:55
    and transcribe it this is called
  • 00:03:58
    the sigma right or you can represent it
  • 00:04:02
    like this
  • 00:04:03
    subunit or factor if you will okay
  • 00:04:07
    these two components the core enzyme
  • 00:04:09
    which is made up of the two alpha the
  • 00:04:10
    beta and the beta prime and the omega
  • 00:04:12
    sub unit
  • 00:04:13
    as well as the sigma subunit make up the
  • 00:04:15
    entire rna polymerase
  • 00:04:16
    now let me show you for example here
  • 00:04:19
    let's say i represent the core enzyme
  • 00:04:21
    as just this kind of blue circle with
  • 00:04:23
    lines in it
  • 00:04:24
    and then we'll represent the sigma
  • 00:04:26
    subunit as kind of like a pink circle
  • 00:04:27
    with some lines in it right so let's
  • 00:04:29
    imagine here
  • 00:04:30
    we have that core enzyme which we're
  • 00:04:33
    going to represent like this
  • 00:04:34
    and then the other component of it which
  • 00:04:36
    is the sigma subunit which will
  • 00:04:38
    represent
  • 00:04:39
    like this that sigma subunit will then
  • 00:04:43
    bind to the promoter region once it
  • 00:04:45
    binds to the promoter region
  • 00:04:47
    then this core enzyme of the rna
  • 00:04:49
    polymerase will then
  • 00:04:50
    release away from the sigma subunit
  • 00:04:54
    and it'll start moving down
  • 00:04:57
    this dna and as it moves down the dna
  • 00:05:00
    it'll read the dna
  • 00:05:01
    from three to five and synthesize an rna
  • 00:05:04
    strand
  • 00:05:05
    from that which we'll talk about more
  • 00:05:07
    detail later
  • 00:05:09
    from five to three
  • 00:05:12
    so it'll read the dna and make rna
  • 00:05:16
    this rna that we make from in
  • 00:05:18
    prokaryotic cells with the rna
  • 00:05:20
    polymerase holoenzyme
  • 00:05:22
    is very different from eukaryotic cells
  • 00:05:25
    in prokaryotic cells that mrna that we
  • 00:05:27
    made from this one rna polymerase
  • 00:05:29
    holoenzyme can make all the mr
  • 00:05:31
    all the rna we need whether that be rrna
  • 00:05:34
    within the prokaryotic cell whether that
  • 00:05:36
    be m
  • 00:05:37
    rna within the prokaryotic cell or t
  • 00:05:41
    rna within the prokaryotic cells
  • 00:05:44
    so that's very important big thing i
  • 00:05:47
    really need you guys to take away from
  • 00:05:48
    that is
  • 00:05:48
    prokaryotic cells they use one rna
  • 00:05:52
    polymerase which is called a holoenzyme
  • 00:05:54
    made up of two components a core enzyme
  • 00:05:56
    made of these subunits
  • 00:05:57
    and a sigma subunit the core is what
  • 00:06:00
    reads the dna and makes the rna the
  • 00:06:02
    sigma subunit is what
  • 00:06:04
    binds the rna polymerase to the promoter
  • 00:06:07
    region
  • 00:06:07
    enabling it to transcribe the dna okay
  • 00:06:11
    and whenever you make rna within a
  • 00:06:13
    prokaryotic cell from this rna
  • 00:06:15
    polymerase it makes
  • 00:06:16
    all the rnas within that prokaryotic
  • 00:06:18
    cell in eukaryotic cells it's a little
  • 00:06:20
    bit different
  • 00:06:22
    so let's talk about that let's say here
  • 00:06:25
    we have
  • 00:06:25
    three promoter regions that i want us to
  • 00:06:27
    focus on and this is all within
  • 00:06:30
    eukaryotic cells
  • 00:06:34
    in eukaryotic cells we need
  • 00:06:38
    two different things in order to allow
  • 00:06:40
    for transcription to occur
  • 00:06:42
    and this portion here right and this
  • 00:06:44
    portion of prokaryotic cells we only
  • 00:06:46
    need
  • 00:06:46
    one enzyme which had two different
  • 00:06:48
    components
  • 00:06:49
    within eukaryotic cells each process
  • 00:06:52
    requires
  • 00:06:53
    a particular enzyme an rna polymerase
  • 00:06:56
    and a transcription factor let's let's
  • 00:06:59
    kind of write that down
  • 00:07:00
    so let's say that we take this first
  • 00:07:01
    promoter we want to read this gene this
  • 00:07:05
    portion of the dna
  • 00:07:06
    and make rna and this is the rna that
  • 00:07:08
    we're actually going to
  • 00:07:10
    synthesize right here okay from this
  • 00:07:13
    gene
  • 00:07:14
    a particular enzyme let's represent this
  • 00:07:16
    in blue since we've been kind of doing
  • 00:07:18
    blue here there's going to be a
  • 00:07:19
    particular enzyme which is going to
  • 00:07:21
    read this dna okay
  • 00:07:24
    and make this rna there's a particular
  • 00:07:26
    enzyme what is that enzyme called it's
  • 00:07:27
    called rna polymerase
  • 00:07:30
    but this is the first promoter within
  • 00:07:33
    the eukaryotic cells that we're talking
  • 00:07:34
    about right
  • 00:07:35
    so let's call it rna polymerase one
  • 00:07:39
    rna polymerase one will read the dna and
  • 00:07:42
    make a particular type of rna but in
  • 00:07:44
    order for it to do this
  • 00:07:46
    it needs a special protein that can bind
  • 00:07:49
    to the promoter region which allows for
  • 00:07:51
    the rna polymerase to bind to the dna
  • 00:07:53
    and read the dna
  • 00:07:54
    what is that particular protein that
  • 00:07:57
    protein
  • 00:07:58
    let's represent it here in let's do
  • 00:08:01
    green there's a particular protein
  • 00:08:05
    which will bind here to the rna
  • 00:08:08
    polymerase
  • 00:08:09
    and to the promoter and allow the rna
  • 00:08:11
    polymerase to
  • 00:08:12
    bind to the dna and start moving down
  • 00:08:15
    reading the dna and making this rna
  • 00:08:17
    what is this called this is called a
  • 00:08:19
    transcription
  • 00:08:20
    factor tf and there's many different
  • 00:08:23
    types of transcription factors the
  • 00:08:26
    particular thing that i need you to
  • 00:08:27
    remember for right now
  • 00:08:29
    is that we call these transcription
  • 00:08:31
    factors which are utilized by rna
  • 00:08:33
    polymerases
  • 00:08:34
    within eukaryotic cells we call these
  • 00:08:38
    general
  • 00:08:40
    transcription factors we'll talk about
  • 00:08:41
    very specific types
  • 00:08:43
    with an rna polymerase type 2 a little
  • 00:08:45
    bit later but for right now
  • 00:08:47
    two things i need in order for this rna
  • 00:08:49
    polymerase to be able to read the dna
  • 00:08:51
    and make this rna rna polymerase 1 needs
  • 00:08:54
    a general transcription factor
  • 00:08:56
    to bind to the promoter allowing the rna
  • 00:08:58
    polymerase 1 to
  • 00:09:00
    then bind into the dna read it and make
  • 00:09:02
    rna
  • 00:09:03
    what type of rna does it make i have all
  • 00:09:06
    the rnas within prokaryotic cells from
  • 00:09:08
    one rna polymerase
  • 00:09:09
    but rna polymerase 1 makes a very
  • 00:09:12
    particular type of rna
  • 00:09:14
    and this is called r rna
  • 00:09:18
    now rrna is very important because this
  • 00:09:21
    is incorporated into what's called
  • 00:09:23
    ribosomes ribosomes and ribosomes are
  • 00:09:27
    utilized in the translation process
  • 00:09:29
    where we take
  • 00:09:30
    mrna and from that make proteins
  • 00:09:34
    so we'll talk about this later in
  • 00:09:35
    another video but for right now first
  • 00:09:36
    thing i need you know is rna polymerase
  • 00:09:38
    one
  • 00:09:38
    with transcription factors reads the dna
  • 00:09:41
    and makes
  • 00:09:42
    rrna now that makes everything else
  • 00:09:44
    pretty easy from this point
  • 00:09:46
    here's another promoter region of a
  • 00:09:48
    particular sequence of dna
  • 00:09:50
    right within a eukaryotic cell so this
  • 00:09:53
    is the second promoter region
  • 00:09:54
    another enzyme binds another rna
  • 00:09:57
    polymerase
  • 00:09:58
    and not only just that one rna
  • 00:10:00
    polymerase here but we also need
  • 00:10:02
    a set of general transcription factors
  • 00:10:06
    to bind to this promoter region so
  • 00:10:09
    general transcription factors we need to
  • 00:10:11
    bind to the promoter region
  • 00:10:12
    enabling this rna polymerase to bind to
  • 00:10:15
    the dna
  • 00:10:15
    read it and then make what make these
  • 00:10:19
    particular types of rna we have here
  • 00:10:23
    this is well this was the first promoter
  • 00:10:26
    this is the second one
  • 00:10:27
    all right so we're going to call this
  • 00:10:29
    rna polymerase
  • 00:10:31
    2 rna polymerase 2 will bind to this
  • 00:10:35
    promoter via the transcription factor
  • 00:10:37
    read the dna and make rna what kind of
  • 00:10:39
    rna is it going to be making
  • 00:10:41
    big thing i need you to remember is it's
  • 00:10:43
    making m
  • 00:10:45
    rna mrna you'll see later again
  • 00:10:48
    is the component it'll have to go
  • 00:10:50
    through some very specific modifications
  • 00:10:54
    that we'll talk about in great detail
  • 00:10:56
    and then eventually
  • 00:10:57
    it'll be translated with
  • 00:11:00
    the help of rrna and another thing
  • 00:11:02
    called trna
  • 00:11:04
    at the ribosomes and making proteins
  • 00:11:06
    okay the other thing that
  • 00:11:08
    you guys can remember if you guys want
  • 00:11:09
    to be scholarly or ninja nerdy
  • 00:11:11
    there's another rna that's made here and
  • 00:11:13
    we'll talk about it a little bit later
  • 00:11:15
    with what's called splicing
  • 00:11:17
    and these are called small nuclear rnas
  • 00:11:21
    and these are involved in what's called
  • 00:11:23
    splicing and we'll get into that a
  • 00:11:25
    little bit more in detail later okay
  • 00:11:27
    but big thing rna polymerase ii with the
  • 00:11:29
    help of general transcription factors
  • 00:11:31
    makes mrna
  • 00:11:32
    and snrnas rna polymerase one with the
  • 00:11:34
    help of general transcription factors
  • 00:11:36
    makes
  • 00:11:36
    rnas when the heck do you think this
  • 00:11:38
    last promoter region of this
  • 00:11:40
    sequence of dna within this eukaryotic
  • 00:11:42
    cell is going to make
  • 00:11:44
    trnas and it's the same process what do
  • 00:11:46
    i need here
  • 00:11:47
    i need general transcription factors to
  • 00:11:49
    bind to the promoter region
  • 00:11:51
    when that binds that facilitates or it
  • 00:11:53
    helps to
  • 00:11:54
    allow the rna polymerase
  • 00:11:57
    type what
  • 00:12:00
    three two
  • 00:12:04
    bind to the dna and then read the dna
  • 00:12:07
    and then synthesize what rna
  • 00:12:12
    what type of rna is it making the type
  • 00:12:14
    of rna that is being synthesized from
  • 00:12:16
    rna polymerase iii is
  • 00:12:18
    primarily trna but
  • 00:12:22
    a teensy little bit of snra is
  • 00:12:25
    also made by rna polymerase type 3.
  • 00:12:28
    and if you guys really want to be extra
  • 00:12:30
    ninja nerdy technically
  • 00:12:32
    even a teensy bit of
  • 00:12:36
    rna is also made here as well okay
  • 00:12:39
    now trna what the heck does this do
  • 00:12:41
    you'll see later
  • 00:12:42
    that this is also involved in the
  • 00:12:44
    translation process
  • 00:12:46
    it carries a particular amino acid and
  • 00:12:48
    an anticodon
  • 00:12:50
    which is going to be involved in that
  • 00:12:51
    process and we'll talk about that in a
  • 00:12:52
    separate video
  • 00:12:53
    so i know this was a lot of stuff to
  • 00:12:54
    take away and take away from this but
  • 00:12:56
    the big
  • 00:12:56
    overall theme that i really just out of
  • 00:12:58
    all of this what i want you to take away
  • 00:13:00
    from this is this quick little thing
  • 00:13:01
    here
  • 00:13:02
    that rna polymerases 1
  • 00:13:06
    2 3 remember
  • 00:13:10
    r m t
  • 00:13:14
    rna polymerase 1 primarily gives way to
  • 00:13:16
    rrna
  • 00:13:17
    rna polymerase 2 primarily gives way to
  • 00:13:20
    mrna
  • 00:13:22
    and then rna polymerase 3 primarily
  • 00:13:24
    gives way to
  • 00:13:25
    t rna these are the big things that i
  • 00:13:28
    want you to take away from all this if
  • 00:13:29
    you want to go the extra mile be extra
  • 00:13:31
    ninja nerdy
  • 00:13:32
    two and 3 also can give way to what
  • 00:13:36
    small nuclear rna if you really want to
  • 00:13:37
    go the extra mile technically three can
  • 00:13:39
    also give way to
  • 00:13:40
    rrna but this is the basic thing to take
  • 00:13:42
    away from what we just talked about
  • 00:13:45
    and then the other thing is in
  • 00:13:47
    prokaryotic cells we don't need all of
  • 00:13:48
    these
  • 00:13:49
    we need one rna polymerase holoenzyme
  • 00:13:53
    to make all the rnas one last thing is
  • 00:13:57
    you notice in eukaryotic cells that we
  • 00:13:58
    have particular transcription factors
  • 00:14:00
    that are going to be needed for each rna
  • 00:14:02
    polymerase
  • 00:14:03
    the transcription factor in prokaryotes
  • 00:14:05
    technically if you want to be specific
  • 00:14:08
    is the sigma subunit because it's the
  • 00:14:09
    portion that's binding to the promoter
  • 00:14:11
    to allow
  • 00:14:12
    the core enzyme of the rna polymerase to
  • 00:14:14
    read the dna
  • 00:14:15
    okay so that kind of covers the basic
  • 00:14:17
    concepts of the two main things that we
  • 00:14:18
    need
  • 00:14:19
    in order for this transcription process
  • 00:14:21
    to occur now
  • 00:14:23
    there's one other thing that i want to
  • 00:14:24
    talk about very quickly before we really
  • 00:14:26
    start talking about
  • 00:14:27
    mrna because that's going to be the
  • 00:14:28
    primary topic here i want to have a
  • 00:14:31
    quick little discussion on how we can
  • 00:14:33
    modulate
  • 00:14:34
    the rate of transcription either
  • 00:14:36
    speeding it up or slowing it down
  • 00:14:38
    okay so the next thing i want to talk
  • 00:14:39
    about is very very briefly
  • 00:14:41
    on eukaryotic gene regulation so i want
  • 00:14:44
    to have a quick
  • 00:14:45
    quick tiny little discussion on gene
  • 00:14:46
    regulation
  • 00:14:48
    okay and the only reason i want to
  • 00:14:49
    mention this is because this is very
  • 00:14:51
    easy and it kind of makes sense along
  • 00:14:53
    with what we're talking about
  • 00:14:55
    but we're not going to talk about it in
  • 00:14:56
    prokaryotic cells we're primarily going
  • 00:14:58
    to talk about this gene regulation
  • 00:15:00
    and eukaryotic cells we're going to have
  • 00:15:02
    a separate video because it's more
  • 00:15:03
    involved
  • 00:15:04
    we'll talk about gene regulation and
  • 00:15:06
    prokaryotic cells with the lac operon
  • 00:15:07
    and the tryptophan operon we'll get into
  • 00:15:09
    that
  • 00:15:09
    but in eukaryotic cells there's two ways
  • 00:15:12
    that we can modulate and it's really
  • 00:15:13
    easy
  • 00:15:14
    one way that we can modulate
  • 00:15:17
    transcription is we have particular dna
  • 00:15:19
    sequences
  • 00:15:20
    particular sequences of dna particularly
  • 00:15:22
    palindromic sequences
  • 00:15:24
    which are called enhancers and enhancers
  • 00:15:28
    are basically dna sequences and
  • 00:15:30
    the big thing i want you to take away
  • 00:15:32
    from this they can increase
  • 00:15:34
    the transcription rate so they increase
  • 00:15:37
    the
  • 00:15:38
    rate of transcription or the process of
  • 00:15:39
    transcription okay we'll talk about how
  • 00:15:41
    they do that
  • 00:15:43
    the other thing that can regulate the
  • 00:15:45
    the transcription process or gene
  • 00:15:47
    regulation in a way
  • 00:15:48
    is something called silencers
  • 00:15:52
    in silencers they do what they decrease
  • 00:15:55
    the transcription rate or the
  • 00:15:57
    transcription process
  • 00:15:59
    now it's really straightforward it's
  • 00:16:02
    relatively simple let me explain
  • 00:16:04
    what i mean let's say here we have a
  • 00:16:06
    strip of dna we're going to explain how
  • 00:16:08
    this happens so here's our strip of dna
  • 00:16:10
    and remember this blue region what did
  • 00:16:12
    we call this blue region that we talked
  • 00:16:13
    about above
  • 00:16:14
    this was called our promoter region and
  • 00:16:17
    do you guys remember let's take
  • 00:16:19
    eukaryotic cells in this case what we
  • 00:16:21
    needed in order for this process to
  • 00:16:24
    occur
  • 00:16:25
    we needed a particular transcription
  • 00:16:27
    factor to bind to that promoter region
  • 00:16:29
    and then what else did we need in order
  • 00:16:31
    for that to read the dna and make rna
  • 00:16:33
    you needed a particular rna polymerase
  • 00:16:37
    right so we need an
  • 00:16:38
    rna polymerase depending on which one
  • 00:16:40
    we're talking about
  • 00:16:41
    would depend on the type of rna that we
  • 00:16:43
    want to make
  • 00:16:44
    and then a transcription factor okay
  • 00:16:48
    now this is going to go read the dna
  • 00:16:50
    this rna polymerase will read the dna
  • 00:16:52
    and then make rna right now here let's
  • 00:16:55
    say that we have the promoter
  • 00:16:56
    and you can have this enhancer upstream
  • 00:17:00
    from the promoter or it could be down
  • 00:17:02
    here downstream where we can't see it in
  • 00:17:03
    this
  • 00:17:04
    diagram but it would be all the way down
  • 00:17:05
    here regardless of where it is it's
  • 00:17:08
    usually can be close to the promoter or
  • 00:17:10
    it can be far to the promoter so you're
  • 00:17:11
    probably asking the question how the
  • 00:17:13
    heck would an enhancer that's really far
  • 00:17:15
    away
  • 00:17:16
    influence a promoter that's all the way
  • 00:17:18
    down here how that does it do that
  • 00:17:20
    there's particular structures there's
  • 00:17:22
    different things
  • 00:17:24
    that can activate enhancers and cause
  • 00:17:27
    conformational changes of the dna
  • 00:17:30
    and these are called specific
  • 00:17:32
    transcription factors you know why i
  • 00:17:34
    really frustrate i got
  • 00:17:35
    really deep into talking about specific
  • 00:17:37
    and general transcription factors
  • 00:17:39
    the general transcription factors are
  • 00:17:40
    what bind to the promoter region
  • 00:17:42
    specific transcription factors which
  • 00:17:45
    we're going to really kind of do a
  • 00:17:46
    different color here let's do purple
  • 00:17:48
    specific transcription factors
  • 00:17:51
    will bind to this enhancer region so
  • 00:17:54
    this let's put specific
  • 00:17:56
    transcription factors these will bind
  • 00:17:59
    to the enhancer when they bind
  • 00:18:02
    to the enhancer region it causes a
  • 00:18:06
    looping of the dna to where now the
  • 00:18:09
    promoter was far
  • 00:18:10
    downstream from this enhancer but when
  • 00:18:13
    this
  • 00:18:14
    specific transcription factor binds to
  • 00:18:16
    the enhancer
  • 00:18:18
    it causes the dna to loop in a way that
  • 00:18:20
    it's in very close proximity to the
  • 00:18:23
    promoter region even though it's far
  • 00:18:24
    upstream from it
  • 00:18:26
    and then what was bound to this promoter
  • 00:18:28
    region here do you guys remember
  • 00:18:31
    the general transcription factors
  • 00:18:34
    and what else the rna
  • 00:18:38
    polymerase so now that these are in
  • 00:18:42
    close proximity guess what this
  • 00:18:44
    general trans this uh specific
  • 00:18:46
    transcription factor can do to this area
  • 00:18:48
    over here
  • 00:18:49
    it can act on these proteins and
  • 00:18:53
    stimulate this reading of the dna
  • 00:18:56
    the rna polymerase is to read the dna
  • 00:18:58
    and to do what
  • 00:19:00
    make rna
  • 00:19:04
    whether it be mrna rrna trna so the
  • 00:19:06
    whole point here is that
  • 00:19:08
    enhancers can be either far upstream or
  • 00:19:10
    far downstream which makes it hard to
  • 00:19:12
    interact with the promoter
  • 00:19:13
    but if a specific transcription factor
  • 00:19:15
    binds to that enhancer it creates a
  • 00:19:17
    looping process
  • 00:19:18
    bringing it in close proximity which can
  • 00:19:21
    then stimulate the
  • 00:19:22
    specific transcription factors and the
  • 00:19:24
    rna polymerases which are bound to the
  • 00:19:26
    promoter
  • 00:19:27
    to increase the transcription of rna
  • 00:19:30
    what do you think silencers do
  • 00:19:32
    the exact same process we're not going
  • 00:19:34
    to go into detail of it
  • 00:19:35
    but if you imagine i did the same thing
  • 00:19:37
    i put the silencer here
  • 00:19:39
    and i have a specific transcription
  • 00:19:41
    factor that bound here it's going to
  • 00:19:42
    fold it in a particular way bringing it
  • 00:19:44
    close to the promoter
  • 00:19:45
    inhibiting that promoter region and
  • 00:19:47
    slowing down the transcription process
  • 00:19:50
    it doesn't make sense it's pretty cool
  • 00:19:52
    too right so i need you guys to ask
  • 00:19:53
    yourself the questions because we're
  • 00:19:54
    going to talk about these
  • 00:19:56
    these general transcription factors what
  • 00:19:58
    in the world
  • 00:20:00
    are these specific transcription factors
  • 00:20:02
    and i know that if you guys are the og
  • 00:20:04
    ninjas
  • 00:20:05
    you'll know these processes in and out
  • 00:20:08
    you guys know when we make a protein
  • 00:20:10
    whenever we have like a cell
  • 00:20:12
    signaling response we've talked about
  • 00:20:13
    this a million times here an engineer
  • 00:20:15
    right
  • 00:20:16
    let's say that we take a hormone like
  • 00:20:18
    tsh which stimulates thyroid hormone
  • 00:20:20
    synthesis right tsh will act on a
  • 00:20:23
    particular receptor we call these
  • 00:20:25
    g-protein-coupled receptors right like
  • 00:20:27
    g-stimulatory proteins
  • 00:20:29
    those g-stimulatory proteins will
  • 00:20:31
    activate something called
  • 00:20:33
    cyclic amp cyclic amp will then activate
  • 00:20:37
    something called
  • 00:20:38
    protein kinase a protein kinase a
  • 00:20:43
    depending upon what type of you know
  • 00:20:46
    transcription factor you need in this
  • 00:20:48
    case
  • 00:20:48
    we're going to activate a very specific
  • 00:20:51
    transcription factor
  • 00:20:52
    for making what thyroid hormone
  • 00:20:55
    so some type of thyroid hormone
  • 00:20:57
    transcription factor
  • 00:20:58
    that'll be needed to bind to the
  • 00:21:00
    enhancer change the shape of it
  • 00:21:02
    activate the promoter have the rna
  • 00:21:05
    polymerase
  • 00:21:06
    read the gene that makes what hormone
  • 00:21:09
    thyroid hormone
  • 00:21:10
    and so you'd have this get read you'd
  • 00:21:12
    make an mrna
  • 00:21:13
    that would then get translated and make
  • 00:21:17
    thyroid hormone doesn't that make sense
  • 00:21:20
    how that process occurs
  • 00:21:22
    so we can increase the transcription and
  • 00:21:25
    protein formation of thyroid hormone
  • 00:21:26
    through this process the same thing
  • 00:21:28
    exists
  • 00:21:28
    with steroid hormones if i took for
  • 00:21:31
    example testosterone
  • 00:21:33
    you guys know testosterone right
  • 00:21:35
    testosterone does what
  • 00:21:38
    testosterone will move across
  • 00:21:41
    the cell membrane it'll bind onto a
  • 00:21:44
    intracellular receptor when testosterone
  • 00:21:48
    binds onto the intracellular receptor
  • 00:21:49
    what will that intracellular receptor do
  • 00:21:52
    bind to the enhancer when it binds to
  • 00:21:54
    the enhancer
  • 00:21:55
    loops it brings it close to the promoter
  • 00:21:56
    stimulates the transcription to make
  • 00:21:58
    proteins within muscle so that you can
  • 00:21:59
    get direct
  • 00:22:00
    right that's the whole process of how we
  • 00:22:03
    increase
  • 00:22:04
    transcription and the same thing would
  • 00:22:06
    happen if we wanted to decrease it just
  • 00:22:08
    we would have some type of
  • 00:22:10
    repressing transcription factor binding
  • 00:22:13
    to the silencer
  • 00:22:14
    that would inhibit the transcription
  • 00:22:15
    process so i think we have a pretty good
  • 00:22:17
    idea now of the basic concepts of
  • 00:22:19
    eukaryotic gene regulation
  • 00:22:21
    now spend most of our time talking about
  • 00:22:23
    the transcription particularly of
  • 00:22:26
    mrna all right so when we talk about
  • 00:22:28
    transcription we've had a basic concept
  • 00:22:29
    of it right that we need rna polymerases
  • 00:22:31
    and transcription factors to read the
  • 00:22:32
    dna and make the
  • 00:22:34
    rna but the real one that i want us to
  • 00:22:36
    primarily focus on
  • 00:22:38
    which is primarily important with
  • 00:22:40
    transcription of dna
  • 00:22:41
    is mrna that was the real important one
  • 00:22:45
    now that's in eukaryotic cells with
  • 00:22:46
    utilizing the what
  • 00:22:48
    rna polymerase type 2 in prokaryotic
  • 00:22:50
    cells we would just be using the
  • 00:22:52
    rna polymerase holoenzyme so what i want
  • 00:22:54
    us to do is i want us to go through
  • 00:22:56
    particularly and more d we already have
  • 00:22:57
    a basic concept of how this is going to
  • 00:22:59
    work
  • 00:22:59
    but let's go into the stages of
  • 00:23:01
    transcription particularly for
  • 00:23:03
    mrna within prokaryotic cells in
  • 00:23:05
    eukaryotic cells
  • 00:23:06
    the first stage that is involved here is
  • 00:23:09
    called initiation
  • 00:23:11
    of transcription so the first step that
  • 00:23:13
    we have to talk about is called
  • 00:23:15
    initiation of transcription now this is
  • 00:23:18
    the part that we've pretty much already
  • 00:23:20
    familiarized ourselves with
  • 00:23:22
    okay now within this let's have our two
  • 00:23:26
    cells
  • 00:23:26
    okay that we're going to do initiation
  • 00:23:28
    with we're going to have our prokaryotic
  • 00:23:30
    cells here
  • 00:23:31
    on this left side of the board
  • 00:23:34
    and then over here we're going to have
  • 00:23:37
    the eukaryotic cells here on the right
  • 00:23:40
    side of the board
  • 00:23:41
    what i want us to do is to have kind of
  • 00:23:42
    a comparison a side-by-side comparison
  • 00:23:45
    of the initiation process the first
  • 00:23:48
    thing that we need to know
  • 00:23:49
    is we've talked a little bit about this
  • 00:23:51
    already but this blue region what did we
  • 00:23:53
    call this blue region here again
  • 00:23:55
    this blue region was called the promoter
  • 00:23:58
    region
  • 00:23:59
    now the promoter region i told you is a
  • 00:24:01
    particular kind of like nucleotide
  • 00:24:03
    sequence that is very very specific and
  • 00:24:05
    allows transcription factors and rna
  • 00:24:07
    polymerases to bind to the dna
  • 00:24:09
    it's kind of a signal if you will it's
  • 00:24:10
    like hey here i am come bind to me
  • 00:24:13
    in prokaryotic cells the promoter
  • 00:24:16
    region has particular types of like
  • 00:24:19
    names and just
  • 00:24:20
    weird stuff that they can ask you in
  • 00:24:22
    your exams
  • 00:24:24
    so in the prokaryotic cells they call
  • 00:24:26
    this the third negative 35 region
  • 00:24:29
    which means from the start point at
  • 00:24:31
    which the rna polymerase starts reading
  • 00:24:33
    the dna and making rna
  • 00:24:35
    if you go back 35 nucleotides that's
  • 00:24:37
    kind of the
  • 00:24:38
    point at which the rna polymerases will
  • 00:24:40
    bind
  • 00:24:41
    in prokaryotic cells another one is
  • 00:24:44
    called the negative 10 region
  • 00:24:46
    but they wanted to give this one a name
  • 00:24:48
    so they called it the pribno box
  • 00:24:50
    just meaning that it's negative it's 10
  • 00:24:52
    nucleotides away from that startup
  • 00:24:54
    transcription right
  • 00:24:56
    and then the last one here is called the
  • 00:24:58
    plus one region which is also called the
  • 00:25:01
    transcription
  • 00:25:02
    start site so it's going to be pretty
  • 00:25:03
    much the nucleotide at which you just
  • 00:25:05
    read
  • 00:25:06
    and start making the whole process of
  • 00:25:09
    rna
  • 00:25:10
    so these are the regions that you guys
  • 00:25:12
    need to remember within prokaryotic
  • 00:25:13
    cells these are the kind of
  • 00:25:14
    specific promoter regions and eukaryotic
  • 00:25:18
    cells
  • 00:25:19
    the promoter regions have particular
  • 00:25:23
    nucleotide sequences that we need to be
  • 00:25:24
    aware of
  • 00:25:26
    these are called the top box which means
  • 00:25:28
    that you would have thymine adenine
  • 00:25:29
    thymine adenine
  • 00:25:30
    that would be a particular recognition
  • 00:25:32
    sequence within the promoter and
  • 00:25:33
    eukaryotic cells
  • 00:25:35
    or cat c-a-a-t so cytosine adenine
  • 00:25:39
    adenine thymine
  • 00:25:40
    and the last one is a gc box
  • 00:25:43
    so if there's a tata box a cap box or a
  • 00:25:46
    gc
  • 00:25:47
    box these are identify identifying
  • 00:25:49
    nucleotide areas at which the rna
  • 00:25:51
    polymerase is type 2
  • 00:25:53
    and transcription factors will bind to
  • 00:25:56
    that is the important thing okay now the
  • 00:25:59
    next thing here is
  • 00:26:00
    the polymerases the rna polymerase is
  • 00:26:03
    within prokaryotic cells it's just
  • 00:26:05
    one it's rna polymerase
  • 00:26:09
    holoenzyme right we already kind of
  • 00:26:12
    talked about that with the core enzyme
  • 00:26:14
    two alpha beta beta prime omega and then
  • 00:26:16
    the
  • 00:26:16
    the sigma subunit all of that's needed
  • 00:26:19
    to bind to the promoter region
  • 00:26:21
    and eukaryotic tells us a little bit
  • 00:26:22
    more right we said that we needed two
  • 00:26:24
    things we needed an rna polymerase
  • 00:26:26
    and which what are we making here
  • 00:26:28
    initiation and we're going to say that
  • 00:26:30
    we're trying to make what
  • 00:26:31
    we're trying to make mrna transcription
  • 00:26:34
    so we're doing
  • 00:26:34
    transcription but we're making mrna so
  • 00:26:37
    what was the particular rna polymerase
  • 00:26:39
    1 2 3 r m m is
  • 00:26:44
    for r for the mrna so rna polymerase
  • 00:26:46
    type 2 is one of the things that i need
  • 00:26:48
    the second thing that i need is the
  • 00:26:52
    general transcription factors
  • 00:26:55
    and there's just so many of these that i
  • 00:26:57
    don't know how important and how
  • 00:26:59
    specific we really need to go into these
  • 00:27:01
    i'll give you some of them but i just
  • 00:27:03
    want you to know
  • 00:27:04
    that there's so many different types of
  • 00:27:06
    them the main one if you had to remember
  • 00:27:09
    one specific out of the tons of them i
  • 00:27:12
    want you to remember transcription
  • 00:27:14
    factor
  • 00:27:14
    2 d this is the one that i really want
  • 00:27:17
    you to remember and the reason why is
  • 00:27:20
    this contains a structure called the
  • 00:27:22
    tata binding protein so this
  • 00:27:23
    transcription factor 2d has a particular
  • 00:27:25
    protein portion
  • 00:27:27
    which binds to the promoter region the
  • 00:27:30
    tata box
  • 00:27:31
    but there's many other reasons region
  • 00:27:34
    transcription factors and you can
  • 00:27:36
    remember these by transcription factor
  • 00:27:38
    2 and there can be h there can be e
  • 00:27:41
    there can be f there can be a there can
  • 00:27:43
    be b so there's tons of these dang
  • 00:27:45
    things
  • 00:27:46
    so i don't know how important it really
  • 00:27:47
    is to know that but the main one i want
  • 00:27:49
    you to remember
  • 00:27:50
    is the transcription factor 2d all right
  • 00:27:54
    so these are the things that we need in
  • 00:27:55
    order for initiation to occur
  • 00:27:57
    so let's take for example we're going to
  • 00:27:58
    have on one side
  • 00:28:00
    eukaryotic cells will the eukaryotic
  • 00:28:02
    enzymes will bind
  • 00:28:03
    and on this side the prokaryotic will
  • 00:28:05
    bind right so let's say here we take for
  • 00:28:07
    example we'll make this prokaryotic rna
  • 00:28:09
    polymerase
  • 00:28:11
    we'll make this one blue
  • 00:28:14
    and we'll make the rna polymerase over
  • 00:28:16
    here for the eukaryotic cells just for
  • 00:28:18
    the heck of it
  • 00:28:19
    we'll make it orange okay just so we can
  • 00:28:21
    distinguish the difference between these
  • 00:28:23
    so what will happen this whole rna
  • 00:28:26
    polymerase holoenzyme will do what
  • 00:28:31
    bind to the promoter what will allow it
  • 00:28:32
    to bind what
  • 00:28:34
    subunit of it the sigma subunit and if
  • 00:28:37
    you really wanted to go back
  • 00:28:38
    you guys remember we made that pink
  • 00:28:42
    okay for the eukaryotic cells
  • 00:28:45
    what do we need we need the rna
  • 00:28:47
    polymerase type 2. we said we're going
  • 00:28:48
    to represent that with
  • 00:28:50
    orange so here's going to be the rna
  • 00:28:53
    polymerase
  • 00:28:54
    type 2 and then what else do we need we
  • 00:28:57
    need those
  • 00:28:58
    general transcription factors there's a
  • 00:29:00
    bunch of them but what's the particular
  • 00:29:02
    one that i really want you to remember
  • 00:29:04
    here
  • 00:29:04
    transcription factor 2 d
  • 00:29:08
    which contains the tata binding protein
  • 00:29:10
    so it binds to the tata box which is the
  • 00:29:12
    promoter region in the eukaryotic cells
  • 00:29:14
    then allows the rna polymerase 2 to bind
  • 00:29:17
    to the dna
  • 00:29:18
    now once the rna polymerase is bound to
  • 00:29:20
    the dna it's going to start
  • 00:29:22
    moving down the dna strands reading it
  • 00:29:25
    and making rna
  • 00:29:26
    so we've now started the process of
  • 00:29:28
    transcription that's all that's
  • 00:29:29
    happening here
  • 00:29:31
    the next step is that once we've bound
  • 00:29:34
    had
  • 00:29:34
    this rna so let's write these down here
  • 00:29:36
    for the prokaryotic cell this would be
  • 00:29:38
    the
  • 00:29:38
    we'll put rna polymerase and we'll put h
  • 00:29:42
    for the holoenzyme and for that one up
  • 00:29:45
    here this is going to be
  • 00:29:47
    rna polymerase type 2
  • 00:29:50
    right once this is bound and it's in the
  • 00:29:53
    dna it's going to start
  • 00:29:54
    reading the dna as it reads the dna
  • 00:29:58
    it'll make mrna that process by which it
  • 00:30:01
    does that is called
  • 00:30:03
    elongation so let's write that down now
  • 00:30:06
    so the next step
  • 00:30:07
    is elongation to make
  • 00:30:11
    the mrna now within elongation a couple
  • 00:30:16
    different things happens
  • 00:30:18
    and this is thankfully the same in
  • 00:30:20
    prokaryotic cells and eukaryotic
  • 00:30:22
    cells so thank the lord for that right
  • 00:30:23
    so let's just say that we take
  • 00:30:25
    either one of these rna polymerases
  • 00:30:27
    let's just for the heck of it we'll say
  • 00:30:29
    here's your rna polymerase ii okay
  • 00:30:33
    here's your rna polymerase two and it's
  • 00:30:34
    reading the dna
  • 00:30:36
    the dna we already know has two strands
  • 00:30:39
    we're going to call this top strand here
  • 00:30:42
    this top strand sonogram this top strand
  • 00:30:44
    up here we're going to call this the
  • 00:30:46
    template strand so the template strand
  • 00:30:50
    also sometimes referred to as the
  • 00:30:55
    anti-sense
  • 00:30:57
    strand this strand down here we're going
  • 00:30:59
    to call
  • 00:31:00
    the coding strand
  • 00:31:04
    now when rna polymerases read dna
  • 00:31:07
    the strand that they read is the
  • 00:31:10
    template strand or the antisense strand
  • 00:31:13
    so that's the first thing i really need
  • 00:31:15
    you guys to remember
  • 00:31:16
    is that the rna polymerases
  • 00:31:20
    what strand do they read they read
  • 00:31:24
    we're going to put the template strand
  • 00:31:27
    or also referred to as what else
  • 00:31:30
    the antisense strand and that's the
  • 00:31:32
    strand that they use to make the mrna
  • 00:31:34
    they do not use the coding strand
  • 00:31:37
    so let's kind of put a little asterisk
  • 00:31:38
    here that this is the strand that we're
  • 00:31:39
    gonna read
  • 00:31:40
    now when it reads it it does it in a way
  • 00:31:43
    that you guys if you guys watch our dna
  • 00:31:44
    replication video this should be
  • 00:31:46
    so darn easy let's say here
  • 00:31:50
    this end of the dna is the three
  • 00:31:53
    prime end that means that this end is
  • 00:31:56
    the
  • 00:31:57
    five prime end and remember one strand
  • 00:32:00
    of dna
  • 00:32:01
    on this side should have a complementary
  • 00:32:04
    anti-parallel strand on the other side
  • 00:32:06
    which means that this is the three and
  • 00:32:08
    on here
  • 00:32:08
    this has to be the five end on this side
  • 00:32:11
    and this has to be the
  • 00:32:13
    three end on that side what happens is
  • 00:32:16
    this rna polymerase when it binds into
  • 00:32:18
    the dna
  • 00:32:19
    it does something very interesting it
  • 00:32:21
    binds to the dna through the initiation
  • 00:32:23
    process
  • 00:32:23
    and then opens up the dna who opened up
  • 00:32:27
    the dna before it was that whole
  • 00:32:29
    in replication it was that whole like
  • 00:32:30
    replication complex
  • 00:32:32
    rna polymerase does that so the first
  • 00:32:33
    thing we need to know is that rna
  • 00:32:35
    polymerase does what
  • 00:32:37
    it opens the dna
  • 00:32:40
    now in replication what else happened
  • 00:32:41
    you opened the dna and you had those
  • 00:32:42
    single stranded binding proteins which
  • 00:32:44
    keep it stable and kept it open right
  • 00:32:46
    rna polymerase does that on its own so
  • 00:32:48
    it also
  • 00:32:49
    stabilizes the single
  • 00:32:53
    stranded dna molecules right so it
  • 00:32:55
    stabilizes the single strands
  • 00:32:57
    then what was the enzyme in replication
  • 00:33:00
    that opened up to unwound the dna
  • 00:33:02
    helicase rna polymerase has its
  • 00:33:05
    intrinsic helicase activity so it also
  • 00:33:08
    unwinds the dna
  • 00:33:12
    after it unwinds the dna then it starts
  • 00:33:14
    reading the dna
  • 00:33:15
    so let's say here as it reads the dna in
  • 00:33:18
    this direction
  • 00:33:19
    three to five it'll make mrna
  • 00:33:24
    that'll be going in the opposite
  • 00:33:26
    direction so it's going to read this 3
  • 00:33:28
    all the way to the 5 direction and as it
  • 00:33:31
    does that it starts synthesizing
  • 00:33:33
    mrna right and this mrna
  • 00:33:37
    will be synthesized in what direction
  • 00:33:38
    what will this be this starting point
  • 00:33:41
    the five end and what would be this
  • 00:33:43
    point
  • 00:33:44
    the three end so we know the next thing
  • 00:33:46
    that the rna polymerase does
  • 00:33:48
    whether it be in prokaryotic cells or
  • 00:33:50
    eukaryotic cells
  • 00:33:52
    is it reads the dna
  • 00:33:55
    from three to five
  • 00:33:58
    then it synthesizes
  • 00:34:02
    rna from what direction guys
  • 00:34:07
    five to three very
  • 00:34:10
    very important the last thing that you
  • 00:34:12
    guys should be asking is okay zach you
  • 00:34:14
    also
  • 00:34:15
    said that in replication the dna
  • 00:34:18
    polymerases read the dna
  • 00:34:20
    and then if there was an accident or a
  • 00:34:22
    mistake they would proofread it and then
  • 00:34:24
    cut out the nucleotide
  • 00:34:25
    what about rna polymerases do they do
  • 00:34:26
    that as well because it looks like
  • 00:34:27
    they've done everything that was similar
  • 00:34:29
    in dna replication
  • 00:34:30
    that's the one thing that's
  • 00:34:31
    controversial so the only thing that's
  • 00:34:33
    kind of relatively controversial
  • 00:34:35
    is is there a proof reading function we
  • 00:34:38
    don't really know it's still subject to
  • 00:34:39
    study
  • 00:34:40
    so that's one thing to remember if you
  • 00:34:42
    want to compare this
  • 00:34:43
    the proofreading function is somewhat
  • 00:34:46
    uncertain
  • 00:34:47
    at this point in time all right so we
  • 00:34:50
    have an idea now we've
  • 00:34:51
    read this dna and we've made rna
  • 00:34:54
    i know we talked about this a lot in dna
  • 00:34:56
    replication we're talking about it here
  • 00:34:58
    and sometimes it really can be confusing
  • 00:35:00
    when you're saying
  • 00:35:01
    five end three and i don't i don't i
  • 00:35:03
    don't freaking get what you're talking
  • 00:35:04
    about zach
  • 00:35:04
    so i want to take a quick little second
  • 00:35:06
    and explain what the heck i mean when i
  • 00:35:08
    say it reads it from three
  • 00:35:09
    to five and synthesizes it from five to
  • 00:35:11
    three a diagram i really think will
  • 00:35:13
    clear this up for you
  • 00:35:14
    let's take a second to understand what i
  • 00:35:16
    mean by reading the dna three to five
  • 00:35:18
    and then
  • 00:35:19
    synthesizing it five to three i think
  • 00:35:20
    it's really important to understand that
  • 00:35:22
    so let's say here we have this strand of
  • 00:35:25
    dna so this is
  • 00:35:26
    this is going to be our dna
  • 00:35:30
    template if you will
  • 00:35:33
    okay so this is our dna template on this
  • 00:35:35
    side the blue one
  • 00:35:36
    and then this is going to be the rna
  • 00:35:39
    that we're going to synthesize utilizing
  • 00:35:41
    the rna polymerase type 2 and eukaryotes
  • 00:35:43
    are the rna polymerase hollow enzyme and
  • 00:35:44
    prokaryotes
  • 00:35:46
    now when we're making this rna we have
  • 00:35:48
    to read
  • 00:35:49
    the dna in what direction the three end
  • 00:35:52
    to the five and what is the three and
  • 00:35:54
    you guys remember the video on dna
  • 00:35:55
    structure
  • 00:35:56
    it's the oh so this is going to be the
  • 00:35:59
    three end
  • 00:35:59
    what's the five end it's the phosphate
  • 00:36:02
    group
  • 00:36:02
    so the phosphate group is going to be
  • 00:36:04
    the five and so i have to read this
  • 00:36:06
    starting at the o h portion towards the
  • 00:36:09
    five end where the phosphate is so the
  • 00:36:13
    rna polymerase let's pretend i'm the rna
  • 00:36:15
    polymerase i'm walking right
  • 00:36:16
    to do i find the three prime and i'm
  • 00:36:18
    like oh there it is okay i'm gonna move
  • 00:36:20
    up oh i found the
  • 00:36:21
    three prime five prime let me just fill
  • 00:36:22
    this up oh i feel my nitrogenous base
  • 00:36:25
    the nitrogenous base that it feels is
  • 00:36:27
    adenine so it picks into its little
  • 00:36:29
    satchel of nucleotides it's like okay
  • 00:36:31
    this is adenine
  • 00:36:32
    the complementary base for is thymine uh
  • 00:36:35
    oh no that's not correct because you
  • 00:36:37
    guys know
  • 00:36:38
    that if we're taking dna making rna
  • 00:36:41
    what's the one nucleotide that
  • 00:36:42
    switches from dna and rna adenine is no
  • 00:36:46
    longer complementary to thymine
  • 00:36:48
    in the rna it is uracil
  • 00:36:52
    so the dna the rna polymerase will come
  • 00:36:54
    read find the three end
  • 00:36:56
    read the nucleotide and say oop okay
  • 00:36:58
    this is an adenine
  • 00:36:59
    reach into its satchel of a bunch of
  • 00:37:01
    nucleotides and pull out
  • 00:37:03
    uracil when it pulls that out it then
  • 00:37:06
    puts the nucleotide in a particular
  • 00:37:08
    orientation
  • 00:37:10
    what's the orientation we said it reads
  • 00:37:11
    it from three to five
  • 00:37:13
    and synthesizes it from five to three
  • 00:37:15
    what's the five
  • 00:37:16
    end here's the nucleotide the five end
  • 00:37:19
    is this
  • 00:37:19
    phosphate group the three end
  • 00:37:22
    is this oh group so it's going to kind
  • 00:37:24
    of flip the nucleotide the opposite
  • 00:37:27
    direction
  • 00:37:28
    and make sure that the nitrogenous base
  • 00:37:30
    here is what
  • 00:37:31
    uracil then when it does that it's going
  • 00:37:34
    to go to the next one so it's going to
  • 00:37:35
    continue it's going to go to the next
  • 00:37:36
    point
  • 00:37:37
    here's where the next oh group would be
  • 00:37:38
    right the three prime end
  • 00:37:40
    reads it finds that finds the nucleotide
  • 00:37:42
    it says oh the nitrogenous base here is
  • 00:37:45
    t let me reach into my satchel of a
  • 00:37:47
    bunch of different uh
  • 00:37:49
    good old nucleotides i'm going to read
  • 00:37:50
    it t goes with a
  • 00:37:52
    i'm going to put my nucleotide and i'm
  • 00:37:53
    going to flip it where it's five prime
  • 00:37:55
    end
  • 00:37:56
    going down three prime end pointing up
  • 00:37:59
    and then the nitrogenous base which is
  • 00:38:02
    complementary to the t
  • 00:38:03
    is a when it does that
  • 00:38:06
    it then fuses the three prime end
  • 00:38:10
    and the five prime end together making a
  • 00:38:13
    bond what is that bond called
  • 00:38:14
    the phosphodiester bond and the same
  • 00:38:17
    process occurs
  • 00:38:18
    so then it'll do what let's fix this
  • 00:38:21
    three prime in there
  • 00:38:22
    it'll then go go to the next nucleotide
  • 00:38:24
    here's the three prime end where the oh
  • 00:38:26
    group is
  • 00:38:26
    reads it finds the nucleotide says that
  • 00:38:29
    it's a g reaches into its satchel pulls
  • 00:38:31
    out a nucleotide
  • 00:38:32
    with the cytosine when it does it it
  • 00:38:34
    flips it to where the five
  • 00:38:35
    end is on this side there's my phosphate
  • 00:38:38
    the three prime end is upwards
  • 00:38:40
    and it says oh the nucleotide that goes
  • 00:38:42
    with this
  • 00:38:43
    is with the nitrogenous base c then it
  • 00:38:46
    says oh
  • 00:38:46
    i have my phi prime n situated close to
  • 00:38:49
    the three prime end of the preceding
  • 00:38:51
    nucleotide
  • 00:38:52
    let me fuse these two together and make
  • 00:38:54
    my phosphate ester bond
  • 00:38:56
    and just for the heck of it because
  • 00:38:57
    repet repetition i guess is helpful
  • 00:38:59
    we go reads this says okay next one
  • 00:39:02
    here's my three prime end where the oh
  • 00:39:03
    group is
  • 00:39:04
    read it find the nitrogenous base it's a
  • 00:39:06
    cytosine
  • 00:39:09
    digs into its satchel pulls out the
  • 00:39:10
    nucleotide guanosine
  • 00:39:13
    sorry the guanine nitrogenous base then
  • 00:39:15
    when it does that
  • 00:39:16
    it situates it where the five prime end
  • 00:39:19
    is situated down
  • 00:39:21
    three prime n is situated upwards in
  • 00:39:22
    this case and then
  • 00:39:24
    the nitrogenous bases on guanine
  • 00:39:28
    then it says oh my five prime n i can
  • 00:39:30
    stitch it to the three prime end of the
  • 00:39:32
    preceding nucleotide
  • 00:39:33
    and form my phosphodiester bond and
  • 00:39:35
    that's how we make rna
  • 00:39:37
    reading it three to five and
  • 00:39:40
    synthesizing it from five to
  • 00:39:42
    three dang we good all right
  • 00:39:45
    now that we've done that the last thing
  • 00:39:47
    i need you to understand is that
  • 00:39:48
    rna polymerase is a very important
  • 00:39:50
    enzyme within eukaryotic and prokaryotic
  • 00:39:52
    cells
  • 00:39:54
    a question that can come up and it's so
  • 00:39:56
    dumb and annoying but you should know it
  • 00:39:59
    is that in eukaryotic cells
  • 00:40:02
    we can inhibit the rna polymerase
  • 00:40:06
    by using a kind of toxin amanitin it's
  • 00:40:10
    for mushrooms
  • 00:40:11
    and this can inhibit the rna polymerase
  • 00:40:14
    within we'll put eukaryotic cells
  • 00:40:18
    okay there's another drug
  • 00:40:21
    which they love to ask in the exams as
  • 00:40:23
    well called rifampicin it's an
  • 00:40:24
    antibiotic
  • 00:40:27
    and this inhibits the rna polymerase
  • 00:40:31
    within if it's an antibiotic that's good
  • 00:40:33
    against bacteria prokaryotic cells
  • 00:40:35
    so this will inhibit the rna polymerase
  • 00:40:37
    within prokaryotic cells
  • 00:40:39
    which would inhibit what the part of the
  • 00:40:42
    initiation
  • 00:40:43
    the elongation basically making rna if
  • 00:40:45
    you can't make rna you can't make
  • 00:40:46
    proteins
  • 00:40:47
    if you can't make proteins you can't
  • 00:40:48
    perform the general functions of the
  • 00:40:50
    cell
  • 00:40:51
    so this is kind of from a poisonous
  • 00:40:54
    mushroom which is stupid to know that
  • 00:40:55
    but they like to ask it on your exams
  • 00:40:57
    and then rifampicin is an antibiotic
  • 00:40:58
    which they also love to ask
  • 00:41:00
    okay now we've talked about elongation
  • 00:41:03
    we've made the dang rna
  • 00:41:05
    rna polymerase is working real hard the
  • 00:41:08
    last thing we got to do is we got to
  • 00:41:09
    just
  • 00:41:09
    end it we don't need any more rna we've
  • 00:41:11
    made the rna that we need to make the
  • 00:41:12
    protein
  • 00:41:13
    that is called termination all right so
  • 00:41:16
    we talked about elongation the next step
  • 00:41:18
    the last step really that we got to
  • 00:41:20
    discuss here is
  • 00:41:21
    termination we've got to end this whole
  • 00:41:23
    transcription process so the last step
  • 00:41:25
    is
  • 00:41:25
    termination now unfortunately
  • 00:41:27
    termination is probably one of the more
  • 00:41:29
    annoying and complicated ones
  • 00:41:31
    unfortunately
  • 00:41:32
    and it is different in prokaryotes and
  • 00:41:34
    eukaryotes that's why it kind of makes
  • 00:41:35
    it a little bit frustrating
  • 00:41:37
    but termination is basically where we've
  • 00:41:40
    already made our rna transcript and we
  • 00:41:43
    just need to
  • 00:41:44
    detach it or disassociate it away from
  • 00:41:46
    the dna and
  • 00:41:47
    prevent that rna polymerase from reading
  • 00:41:50
    any more of the dna and making any more
  • 00:41:51
    rna so just stop
  • 00:41:52
    transcription how do we do that in
  • 00:41:55
    prokaryotes there's two mechanisms
  • 00:41:58
    one of the ways that this happens is
  • 00:42:00
    through what's called
  • 00:42:02
    road dependent termination so one is via
  • 00:42:04
    this process called row
  • 00:42:07
    dependent termination and it's really
  • 00:42:10
    simple believe it or not
  • 00:42:12
    so let's say here we take the
  • 00:42:13
    prokaryotics we we picked blue for our
  • 00:42:16
    rna polymerase so the rna polymerase
  • 00:42:18
    here's our rna polymerase
  • 00:42:20
    it's reading this dna as it's reading
  • 00:42:23
    the dna again what is it making from it
  • 00:42:26
    you guys remember it's making the rna in
  • 00:42:29
    this case
  • 00:42:30
    it could be any rna it could be the mrna
  • 00:42:32
    trna rna whatever
  • 00:42:35
    as it does this there's a protein called
  • 00:42:37
    rho
  • 00:42:38
    and what rho does is this rho protein
  • 00:42:42
    will start moving up the mrna
  • 00:42:45
    and as it moves up the rna that's being
  • 00:42:47
    synthesized by the rna polymerase as it
  • 00:42:49
    gets to this rna polymerase it basically
  • 00:42:53
    says hey
  • 00:42:54
    it just punches the rna polymerase off
  • 00:42:56
    the dna
  • 00:42:58
    if you punch the rna polymerase off the
  • 00:43:01
    dna
  • 00:43:02
    is it going to be able to continue to
  • 00:43:03
    keep breeding the dna and making
  • 00:43:05
    any more rna no so that terminates the
  • 00:43:08
    transcription process
  • 00:43:10
    so the big thing i need you guys to know
  • 00:43:11
    here is that with the road dependent
  • 00:43:13
    termination
  • 00:43:14
    is rho protein
  • 00:43:17
    causes rna polymerase
  • 00:43:21
    uh to break away to disassociate if you
  • 00:43:25
    will
  • 00:43:25
    okay to break
  • 00:43:29
    away from the dna okay
  • 00:43:32
    all right beautiful the next mechanism
  • 00:43:35
    within prokaryotes
  • 00:43:37
    is rho independent termination so we
  • 00:43:40
    don't use the row protein
  • 00:43:41
    so we call this row
  • 00:43:45
    independent termination now with this
  • 00:43:48
    process it's a little bit more
  • 00:43:49
    complicated and a little annoying
  • 00:43:52
    let's say here we have the dna right and
  • 00:43:54
    within the dna we're going to mark these
  • 00:43:56
    here we're going to say this is our
  • 00:43:57
    template strand right so this strand is
  • 00:43:59
    the template strand
  • 00:44:01
    right or the antisense strand and then
  • 00:44:03
    this is going to be our coding strand
  • 00:44:05
    so which one does the rna polymerase
  • 00:44:08
    read it reads the
  • 00:44:09
    template strand or the antisense strand
  • 00:44:12
    there's a particular like thing called
  • 00:44:14
    inverted repeats that form within the
  • 00:44:17
    dna that the rna polymerase is reading
  • 00:44:20
    so what happens is this rna polymerase
  • 00:44:22
    will bind
  • 00:44:23
    to that template strand and it'll start
  • 00:44:25
    reading it
  • 00:44:26
    making the rna as it starts making this
  • 00:44:29
    rna
  • 00:44:30
    it it encounters a particular sequence
  • 00:44:33
    of
  • 00:44:34
    of dna called inverted repeats let's
  • 00:44:37
    write these inverted repeats out in kind
  • 00:44:38
    of a nice little color let's do let's do
  • 00:44:40
    orange
  • 00:44:42
    and let's say here we have an inverted
  • 00:44:43
    repeat where we have c
  • 00:44:45
    c g g and then a bunch of nucleotides
  • 00:44:48
    that we don't care about
  • 00:44:50
    and then here we'll have ggcc
  • 00:44:55
    okay then we're just going to have this
  • 00:44:57
    is the template again
  • 00:44:58
    on the coding strain it would just be
  • 00:45:00
    the complementary base so if this was cc
  • 00:45:02
    this would be gg
  • 00:45:04
    cc we don't really care about these
  • 00:45:06
    nucleotides cc
  • 00:45:08
    gg right the rna polymerase is going to
  • 00:45:11
    read this template strand what happens
  • 00:45:13
    is right you're going to get this kind
  • 00:45:15
    of strand here where you'll have
  • 00:45:17
    a bunch of nucleotides already kind of
  • 00:45:18
    made up here
  • 00:45:20
    and then it reaches this kind of like
  • 00:45:21
    inverted repeat area
  • 00:45:23
    and what happens is it reads this and
  • 00:45:25
    then basically everything you read
  • 00:45:27
    within the template strand should be the
  • 00:45:28
    same as it is in the coding strand
  • 00:45:30
    because it's the complementary base
  • 00:45:31
    so you'll have g g c
  • 00:45:34
    c that it'll make a bunch of nucleotides
  • 00:45:36
    we don't care about
  • 00:45:37
    and then c c g g
  • 00:45:41
    what happens is whenever this
  • 00:45:44
    rna is kind of coming and being
  • 00:45:46
    transcribed from the rna polymerase
  • 00:45:48
    something interesting happens where some
  • 00:45:50
    of these c's and some of these g's on
  • 00:45:53
    this portion have a strong affinity for
  • 00:45:56
    some of the c's and some of the g's
  • 00:45:58
    in this portion of the rna and as they
  • 00:46:01
    start having this affinity they start
  • 00:46:02
    approaching and kind of wanting to
  • 00:46:04
    interact with one another via these
  • 00:46:05
    hydrogen bonds
  • 00:46:07
    and so it creates this really
  • 00:46:09
    interesting kind of like hairpin loop if
  • 00:46:11
    you will
  • 00:46:12
    where there's a bunch of g's and c's
  • 00:46:15
    within this kind of hairpin loop
  • 00:46:19
    that are kind of interacting with one
  • 00:46:21
    another and what happens
  • 00:46:23
    is that hairpin loop is what
  • 00:46:26
    triggers the rna polymerase to pretty
  • 00:46:29
    much
  • 00:46:30
    hop off of the dna and terminate the
  • 00:46:32
    transcription process
  • 00:46:34
    because what happens is once you form
  • 00:46:35
    this hairpin loop what will happen is
  • 00:46:37
    there's going to be particular enzymes
  • 00:46:39
    that will
  • 00:46:39
    bind to that portion and cleave the d
  • 00:46:42
    the rna
  • 00:46:43
    away from the rna polymerase so the big
  • 00:46:46
    thing i need you to
  • 00:46:47
    know within row independent termination
  • 00:46:50
    is that you'll hit this area the rna
  • 00:46:51
    polymerase will be transcribing
  • 00:46:53
    reading the dna making rna it'll hit
  • 00:46:55
    these areas of inverted repeats
  • 00:46:58
    when these inverted repeats are made
  • 00:47:00
    they create this thing called a
  • 00:47:03
    hairpin loop this hairpin loop
  • 00:47:07
    will then trigger particular cleavage
  • 00:47:10
    enzymes
  • 00:47:11
    to come and cleave a couple nucleotides
  • 00:47:15
    after that hairpin loop
  • 00:47:16
    to cleave that away from the rna
  • 00:47:19
    polymerase
  • 00:47:20
    and then here you have your rna that you
  • 00:47:23
    formed
  • 00:47:24
    so that is one of the ways that we have
  • 00:47:25
    termination road
  • 00:47:27
    independent via prokaryotes the last
  • 00:47:30
    termination mechanism is going to be
  • 00:47:32
    eukaryotic cells now how does this work
  • 00:47:36
    this one's actually relatively simple so
  • 00:47:38
    we had the rna polymerase in eukaryotes
  • 00:47:40
    and this was orange
  • 00:47:42
    okay it's binding to the dna it's
  • 00:47:44
    reading the dna
  • 00:47:45
    as it's reading the dna it's making rna
  • 00:47:50
    as it starts making this rna it hits a
  • 00:47:52
    particular sequence
  • 00:47:54
    where when it starts reading the dna and
  • 00:47:56
    makes rna
  • 00:47:58
    it makes a particular sequence of
  • 00:48:01
    a a u a a
  • 00:48:04
    a okay so what are the what is the
  • 00:48:06
    nucleotide sequence here let's write it
  • 00:48:08
    out
  • 00:48:08
    this portion here will be double a u
  • 00:48:11
    triple a this is what's called a
  • 00:48:14
    polyadenylation signal
  • 00:48:16
    so what is this called here this is
  • 00:48:18
    called a poly
  • 00:48:19
    adenylation signal
  • 00:48:24
    and once this kind of nucleotide
  • 00:48:27
    sequence occurs
  • 00:48:28
    so it's kind of now that we know what
  • 00:48:29
    that nucleotide sequence is let's kind
  • 00:48:30
    of just put like this
  • 00:48:33
    here's that nucleotide sequence that
  • 00:48:34
    polyadenylation signal
  • 00:48:36
    that's been synthesized or formed by the
  • 00:48:38
    rna polymerase with the eukaryotes
  • 00:48:40
    once that happens it activates
  • 00:48:42
    particular
  • 00:48:43
    enzymes and those enzymes will come to
  • 00:48:46
    the area here
  • 00:48:49
    and cleave the rna away from the rna
  • 00:48:52
    polymerase
  • 00:48:53
    separating out this rna away
  • 00:48:58
    from the dna and the rna polymerase and
  • 00:49:00
    then again what will i have at this
  • 00:49:01
    portion here
  • 00:49:02
    just as kind of a diagrammatic portion
  • 00:49:05
    here this will be my
  • 00:49:06
    polyadenylation signal this is important
  • 00:49:09
    because we're going to talk about
  • 00:49:10
    post-transcriptional modification in a
  • 00:49:11
    second
  • 00:49:12
    so i know this was a lot of crap just
  • 00:49:14
    really quickly
  • 00:49:15
    recap because this is one of the
  • 00:49:16
    toughest parts of transcription
  • 00:49:18
    is termination prokaryotes there's two
  • 00:49:20
    ways road dependent row independent
  • 00:49:23
    with this one you need a row protein to
  • 00:49:25
    knock the rna polymerase off if you
  • 00:49:26
    don't have him he can't make any more
  • 00:49:28
    rna
  • 00:49:29
    the other one is row independent you
  • 00:49:30
    don't have a row protein
  • 00:49:32
    the rna polymerase is reading the dna
  • 00:49:34
    making rna and it hits these areas of
  • 00:49:37
    inverted repeats these inverted repeats
  • 00:49:40
    when they're made within the
  • 00:49:41
    rna it creates a hydrogen bond
  • 00:49:44
    interaction between them
  • 00:49:46
    which causes it to loop forming a
  • 00:49:47
    hairpin loop that
  • 00:49:49
    signals particular enzymes to break the
  • 00:49:52
    rna
  • 00:49:53
    away from the rna polymerase and we've
  • 00:49:55
    made our rna there
  • 00:49:56
    the last one is in eukaryotes the rna
  • 00:49:59
    polymerase is reading the dna
  • 00:50:01
    and it reaches a particular sequence of
  • 00:50:02
    nucleotides where it reads
  • 00:50:04
    and then makes a a u
  • 00:50:08
    triple a a polyadenylation signal which
  • 00:50:11
    activates enzymes to come
  • 00:50:12
    cleave the rna away from the rna
  • 00:50:14
    polymerase terminating
  • 00:50:16
    the transcription process that really
  • 00:50:19
    hammers this home let's now talk about
  • 00:50:21
    post-transcriptional modification
  • 00:50:22
    we know at this point how to take dna
  • 00:50:25
    make rna
  • 00:50:26
    right we talked about all the different
  • 00:50:27
    types of rna utilizing rna polymerases
  • 00:50:29
    utilizing the transcription factors we
  • 00:50:30
    talked a little about a gene regulation
  • 00:50:32
    we even went through all the stages of
  • 00:50:34
    transcription
  • 00:50:36
    taking the dna and making the mrna
  • 00:50:39
    all the way up until the point where we
  • 00:50:41
    finally made the mrna and broken it away
  • 00:50:44
    from the dna
  • 00:50:45
    unfortunately that's not it for
  • 00:50:47
    transcription
  • 00:50:48
    now we have this mrna right so we
  • 00:50:51
    basically what have we covered up to
  • 00:50:53
    this point
  • 00:50:54
    we took the dna
  • 00:50:57
    we read let's just say here
  • 00:51:01
    at this portion i'll just put here's our
  • 00:51:03
    promoter
  • 00:51:04
    our rna polymerase has read this gene
  • 00:51:06
    sequence
  • 00:51:08
    we hit a termination sequence let's say
  • 00:51:10
    here's our termination sequence
  • 00:51:12
    that we talked about here and once we
  • 00:51:15
    hit that termination sequence
  • 00:51:17
    the rna polymerase will fall off and
  • 00:51:19
    then from this you'll make the
  • 00:51:21
    rna so this was pretty much the basic
  • 00:51:23
    aspects of the transcription
  • 00:51:26
    but now we got to modify this now here's
  • 00:51:28
    the thing
  • 00:51:30
    it's actually kind of a misnomer to say
  • 00:51:32
    that this
  • 00:51:33
    is mrna it's technically not mrna right
  • 00:51:37
    now
  • 00:51:37
    so this piece of rna that we made okay
  • 00:51:41
    and this is this process of
  • 00:51:42
    post-transcriptional modification
  • 00:51:44
    this only occurs it's very important let
  • 00:51:46
    me actually write this down
  • 00:51:47
    this only occurs in
  • 00:51:50
    eukaryotic cells so that's nice
  • 00:51:53
    all this stuff that we're going to talk
  • 00:51:54
    about here is only in eukaryotic cells
  • 00:51:56
    it doesn't happen in prokaryotic cells
  • 00:51:57
    so they just make their rna and that's
  • 00:51:59
    it so technically
  • 00:52:01
    this immature mrna if you will we
  • 00:52:03
    actually give it a very specific name we
  • 00:52:05
    call it heterogeneous
  • 00:52:07
    nuclear rna now
  • 00:52:10
    this heterogeneous nuclear rna is kind
  • 00:52:12
    of an immature
  • 00:52:14
    mrna that has to go through some
  • 00:52:15
    modifications to really make mature
  • 00:52:18
    mrna that then can be translated to make
  • 00:52:20
    proteins
  • 00:52:21
    what are those modifications the first
  • 00:52:24
    thing that we have to do
  • 00:52:26
    is we have to put something on one of
  • 00:52:29
    these ends so now we got to know a
  • 00:52:30
    little bit about the
  • 00:52:31
    terminology of the ends of this immature
  • 00:52:35
    mrna or hn rna on this end
  • 00:52:38
    we're going to call this the five prime
  • 00:52:41
    end what's on that five prime end do you
  • 00:52:42
    guys remember
  • 00:52:43
    the phosphate groups what's on this end
  • 00:52:46
    the three prime end what's on the three
  • 00:52:47
    prime mint
  • 00:52:48
    the oh group okay now
  • 00:52:51
    something very interesting is on the
  • 00:52:52
    five prime end on the five prime end of
  • 00:52:55
    this heterogeneous nuclear rna or the
  • 00:52:57
    hrna
  • 00:52:58
    you have a triphosphate which we're
  • 00:53:00
    representing here with these
  • 00:53:02
    orange circles an enzyme
  • 00:53:05
    comes to the rescue and cleaves off
  • 00:53:09
    one of those phosphate molecules what is
  • 00:53:11
    that enzyme called
  • 00:53:12
    it's this orange little cute enzyme this
  • 00:53:15
    orange enzyme is called
  • 00:53:17
    rna tri-phosphatase
  • 00:53:23
    and what it does is it comes and cleaves
  • 00:53:26
    off
  • 00:53:27
    what portion it cleaves off one of these
  • 00:53:31
    phosphate groups it's going to cleave
  • 00:53:33
    off one of the phosphate groups
  • 00:53:34
    so now i only have two phosphates on the
  • 00:53:37
    end
  • 00:53:38
    of this five prime end then another
  • 00:53:41
    enzyme comes in
  • 00:53:42
    and it says hey there's only two
  • 00:53:44
    phosphates here
  • 00:53:45
    i can now add something on here and i'm
  • 00:53:49
    going to add on what's called a
  • 00:53:52
    gmp molecule what am i going to add on
  • 00:53:54
    again
  • 00:53:55
    i'm going to add a gmp molecule which is
  • 00:53:58
    guanosine monophosphate so we're going
  • 00:54:01
    to represent that here which
  • 00:54:03
    we add on the phosphate for the
  • 00:54:05
    guanosine monophosphate
  • 00:54:07
    and then we're going to just represent
  • 00:54:08
    this as the guanosine so this is our
  • 00:54:10
    guanosine and that blue circle there is
  • 00:54:12
    the phosphate on the guanosine
  • 00:54:14
    so what does he add on technically
  • 00:54:17
    he adds on to this little two phosphates
  • 00:54:20
    right
  • 00:54:21
    it adds in gtp but
  • 00:54:24
    when it does that two phosphates are
  • 00:54:27
    released
  • 00:54:28
    in the form of pyrophosphate which then
  • 00:54:31
    get broken down by pyrophosphatase into
  • 00:54:34
    individual phosphates so if i took gtp
  • 00:54:37
    and i removed two phosphates what am i
  • 00:54:39
    left with
  • 00:54:40
    gmp so it adds on this gmp group
  • 00:54:43
    onto that two phosphate end on the five
  • 00:54:46
    prime end
  • 00:54:47
    so this enzyme that adds that gmp on in
  • 00:54:50
    the form of gtp is called
  • 00:54:52
    guanolile
  • 00:54:55
    uh transferase
  • 00:54:58
    guanalyl transferase beautiful
  • 00:55:02
    so this last enzyme here which is
  • 00:55:03
    involved in this step here on the five
  • 00:55:05
    prime n
  • 00:55:06
    is going to add on a methyl group onto
  • 00:55:09
    one of the
  • 00:55:10
    components of the guanosine
  • 00:55:11
    monophosphate it's actually like one of
  • 00:55:14
    the seventh
  • 00:55:14
    components on that structure it adds on
  • 00:55:17
    a methyl group
  • 00:55:19
    and so at the end of this this enzyme
  • 00:55:23
    which adds a methyl group on what do you
  • 00:55:24
    think it's called methyltransferase
  • 00:55:30
    at the end of this process where you
  • 00:55:32
    took the prime end which had three
  • 00:55:33
    phosphates got rid of one
  • 00:55:36
    took the guanola transfers added on the
  • 00:55:37
    gmp took the methyl transferase added on
  • 00:55:40
    that methyl group
  • 00:55:41
    you formed this complex here and we call
  • 00:55:43
    this whole complex that we just added on
  • 00:55:46
    a seven methyl guanosine group
  • 00:55:49
    okay and that's on that five prime end
  • 00:55:53
    this is called capping
  • 00:55:57
    this is called capping so whatever we've
  • 00:55:59
    just done on this
  • 00:56:00
    five prime end is called capping what
  • 00:56:03
    the heck do we do all this stuff for
  • 00:56:05
    the whole purpose of capping
  • 00:56:08
    is to help to initiate
  • 00:56:13
    translation so this sequence this kind
  • 00:56:15
    of five prime end
  • 00:56:16
    with that seven methyl guanosine or that
  • 00:56:18
    five prime capping if you will
  • 00:56:21
    it's kind of a signal sequence if you
  • 00:56:23
    will that allow for it to interact with
  • 00:56:25
    the ribosome
  • 00:56:26
    and undergo translation the other thing
  • 00:56:28
    it does is it
  • 00:56:30
    prevents degradation by
  • 00:56:33
    nuclease enzymes that want to come and
  • 00:56:35
    break down
  • 00:56:36
    the rna so it helps to prevent
  • 00:56:38
    degradation helps to initiate the
  • 00:56:40
    translation process
  • 00:56:43
    one more thing that they it's a dumb
  • 00:56:45
    thing to know but they love to ask it
  • 00:56:47
    is that there is a particular molecule
  • 00:56:51
    that this methyltransferase
  • 00:56:52
    uses to add that methyl group on and
  • 00:56:55
    sometimes it's really important to know
  • 00:56:56
    it
  • 00:56:57
    and this is called s adenosyl methionine
  • 00:57:00
    also known as
  • 00:57:01
    sam sam carries a methyl group it's like
  • 00:57:04
    a methyl donor if you will
  • 00:57:06
    it gives that methyl group to the methyl
  • 00:57:09
    transferase
  • 00:57:10
    and the methyl transferase adds that
  • 00:57:12
    methyl group onto the guanosine
  • 00:57:13
    monophosphate forming the
  • 00:57:15
    7-methylguanosine
  • 00:57:17
    or that 5-prime cap okay
  • 00:57:20
    so that's the first thing that happens
  • 00:57:22
    now we got to talk about the 3-prime end
  • 00:57:24
    on the three prime end we had that oh
  • 00:57:26
    group right that's the ohn but do you
  • 00:57:28
    remember
  • 00:57:30
    in eukaryote there was a particular
  • 00:57:32
    signal
  • 00:57:33
    that prevent that generated that
  • 00:57:36
    terminated transcription what was that
  • 00:57:38
    nucleotide signal do you guys remember
  • 00:57:40
    test your knowledge guys
  • 00:57:42
    a a u
  • 00:57:45
    triple a right that was that
  • 00:57:47
    polyadenylation signal do you guys
  • 00:57:49
    remember that the polyadenylation
  • 00:57:50
    segment we talked about in eukaryotes
  • 00:57:53
    that polyadenylation signal is
  • 00:57:55
    recognizable
  • 00:57:56
    by this cute little purple enzyme here
  • 00:57:59
    this cute little purple enzyme is called
  • 00:58:01
    poly a
  • 00:58:04
    polymerase it's called
  • 00:58:08
    poly a polymerase what it does is on
  • 00:58:11
    this
  • 00:58:11
    hand it has a bunch of
  • 00:58:15
    adenine
  • 00:58:19
    nucleotides right so it can eat a lot of
  • 00:58:21
    nucleotides containing the adenine
  • 00:58:22
    nitrogenous base
  • 00:58:24
    it takes one end and identifies that
  • 00:58:27
    polyadenylation signal
  • 00:58:28
    takes the other end and adds on
  • 00:58:32
    all of those adenine nucleotides a bunch
  • 00:58:37
    of them
  • 00:58:37
    sometimes up to 200 adenine nucleotides
  • 00:58:41
    when it does that this forms a tail
  • 00:58:44
    on that three prime end with a bunch of
  • 00:58:47
    adenine nucleotides and we call this the
  • 00:58:50
    poly a tail
  • 00:58:54
    so the poly a tail what's the purpose of
  • 00:58:58
    this
  • 00:58:58
    it's the exact same thing helps to
  • 00:59:01
    initiate
  • 00:59:02
    the translation process
  • 00:59:06
    and helps to decrease degradation
  • 00:59:10
    by what kind of enzymes nucleases that
  • 00:59:13
    will try to come and break down that end
  • 00:59:15
    okay the other thing that they do is
  • 00:59:17
    they help transport this
  • 00:59:20
    hn rna eventually they're going to help
  • 00:59:22
    to transport the
  • 00:59:24
    hnrna which will become mrna out of the
  • 00:59:26
    nucleus into the cytosol so they also
  • 00:59:28
    play a little bit of a role in
  • 00:59:29
    transport out of the nucleus and into
  • 00:59:32
    the cytosol
  • 00:59:34
    okay so within this first step what did
  • 00:59:37
    we do
  • 00:59:38
    we did five prime capping we went over
  • 00:59:40
    that part and the three prime
  • 00:59:42
    poly a tail that we did okay now we have
  • 00:59:46
    this so after we did all of this massive
  • 00:59:48
    mess
  • 00:59:49
    we've come to this point
  • 00:59:53
    okay on this part what do we have
  • 00:59:57
    we're just going to write these we're
  • 00:59:58
    going to circle it here this is our
  • 01:00:00
    five prime cap with the
  • 01:00:01
    7-methylguanosine
  • 01:00:03
    and on this end we already have kind of
  • 01:00:05
    formed our
  • 01:00:07
    polyetail the next thing that happens is
  • 01:00:11
    what's called
  • 01:00:12
    splicing and this can be sometimes a
  • 01:00:14
    little annoying
  • 01:00:15
    but it's not too bad i promise let's say
  • 01:00:19
    here
  • 01:00:20
    this is the sequence of nucleotides
  • 01:00:22
    within this
  • 01:00:23
    rna okay we're not at mrna yet we're
  • 01:00:27
    still at this h in rna we're still kind
  • 01:00:29
    of at this h
  • 01:00:30
    in rna at this point we haven't made
  • 01:00:32
    mrna yet
  • 01:00:34
    within this hn rna there's particular
  • 01:00:37
    nucleotides that will be read translated
  • 01:00:41
    and actually will code for particular
  • 01:00:43
    amino acids
  • 01:00:45
    there's other nucleotides within this h
  • 01:00:48
    rna
  • 01:00:48
    that will not be read and they do not
  • 01:00:51
    code for a particular amino acid
  • 01:00:53
    we give those very specific names i'm
  • 01:00:56
    going to highlight them
  • 01:00:57
    in different colors so let's say i
  • 01:01:00
    highlight this one here
  • 01:01:02
    and pink and then i will highlight
  • 01:01:05
    this one here in this kind of maroon
  • 01:01:07
    color
  • 01:01:09
    and then i'll pick here a blue
  • 01:01:13
    and then we'll do another maroon color
  • 01:01:14
    and then we'll do one more color after
  • 01:01:16
    this here's another maroon
  • 01:01:18
    and then we will do just for the heck of
  • 01:01:21
    it black
  • 01:01:23
    okay these portions here
  • 01:01:27
    the pink one this is actually going to
  • 01:01:29
    code
  • 01:01:30
    for an amino acid if it codes for an
  • 01:01:33
    amino acid we give a very specific name
  • 01:01:35
    for that
  • 01:01:36
    and we call it exons so exons
  • 01:01:39
    code for an amino acid
  • 01:01:43
    okay particularly amino acids will make
  • 01:01:46
    proteins
  • 01:01:47
    these other portions and again that's
  • 01:01:49
    going to be or we'll
  • 01:01:51
    i'll mention which ones are exons and
  • 01:01:52
    which ones are the next thing which is
  • 01:01:54
    called
  • 01:01:54
    introns introns are basically
  • 01:01:57
    nucleotide sequences that
  • 01:02:01
    do not code
  • 01:02:05
    for amino acids which will help to make
  • 01:02:08
    proteins
  • 01:02:09
    very important i'm going to call this
  • 01:02:12
    pink portion of the h and rna
  • 01:02:15
    i'm going to call this an exon but we
  • 01:02:16
    have a bunch of them in this h and rna
  • 01:02:18
    so i'm going to call this exon one
  • 01:02:20
    okay that's going to code for some amino
  • 01:02:22
    acids
  • 01:02:24
    i'm going to have this portion here
  • 01:02:25
    which is going to be in the maroon i'm
  • 01:02:26
    going to call this
  • 01:02:27
    an intron but you can have multiple
  • 01:02:29
    introns so i'm going to call this intron
  • 01:02:31
    1.
  • 01:02:32
    same thing here this is going to be
  • 01:02:34
    coding
  • 01:02:35
    so if it codes it's what it's an exon
  • 01:02:38
    well we have multiple types so we're
  • 01:02:39
    going to call this exon 2.
  • 01:02:42
    then i'm going to test you again this
  • 01:02:44
    one does not code for amino acids
  • 01:02:46
    so this is going to be a intron but we
  • 01:02:49
    have already intron once we're going to
  • 01:02:50
    call this
  • 01:02:51
    intron 2 and you guys already kind of
  • 01:02:52
    get the the pattern that i'm going with
  • 01:02:54
    here
  • 01:02:55
    this one does code so it's going to be a
  • 01:02:58
    exon
  • 01:02:59
    and we already have 1 2 so this will be
  • 01:03:01
    three
  • 01:03:02
    okay we're going to do something called
  • 01:03:05
    splicing
  • 01:03:06
    where let's think about this if the
  • 01:03:09
    introns don't
  • 01:03:10
    code for any amino acids do we even need
  • 01:03:12
    them
  • 01:03:13
    no let's get rid of them that's all that
  • 01:03:15
    splicing is
  • 01:03:16
    it's getting rid of these introns or
  • 01:03:18
    also known as intervening sequences
  • 01:03:21
    and then stitching together the exons
  • 01:03:25
    now in order for that process to occur
  • 01:03:29
    we need very specific molecules and we
  • 01:03:32
    talked about it before
  • 01:03:33
    let's see if you guys remember him rna
  • 01:03:36
    polymerase two and three they made
  • 01:03:38
    another very interesting small little
  • 01:03:39
    rna what was that rna called
  • 01:03:42
    small nuclear rna right
  • 01:03:45
    original right so small nuclear rna
  • 01:03:48
    is gonna combine we haven't used this
  • 01:03:50
    color yet so let's add this
  • 01:03:53
    these brown proteins okay so you're
  • 01:03:56
    gonna have some proteins
  • 01:03:57
    and some small nuclear rna together
  • 01:04:01
    these two things make up a very weird
  • 01:04:04
    name
  • 01:04:05
    called a snurp
  • 01:04:08
    okay snurps small nuclear
  • 01:04:12
    ribo ribonuclear proteins so our small
  • 01:04:14
    nuclear ribonuclear proteins and what
  • 01:04:16
    they do
  • 01:04:16
    is these snurps are going to bind
  • 01:04:21
    to this hn rna and they're going to
  • 01:04:24
    cleave
  • 01:04:25
    out the introns in this this actual rna
  • 01:04:29
    and then they're going to stitch
  • 01:04:30
    together the exons
  • 01:04:32
    so let's show that in a very basic way
  • 01:04:34
    of how that happens
  • 01:04:35
    so these snares which are the snra and
  • 01:04:38
    your proteins
  • 01:04:40
    are going to perform splicing so what
  • 01:04:44
    would that look like let's let's take
  • 01:04:45
    here
  • 01:04:47
    our transcript here and bring it down
  • 01:04:49
    here all the way down
  • 01:04:50
    to this portion here
  • 01:04:54
    so here we're going to make our
  • 01:04:55
    functional mrna so at this point in time
  • 01:04:57
    we've actually made what
  • 01:04:59
    at this point we've made the mature
  • 01:05:02
    mrna and
  • 01:05:05
    if i were to show kind of what was the
  • 01:05:09
    end result what am i going to have here
  • 01:05:11
    let's say here i have a sequence that's
  • 01:05:12
    in pink that's exon one
  • 01:05:15
    i got rid of intron one so what should
  • 01:05:17
    be next
  • 01:05:18
    i should have exon two
  • 01:05:22
    i got rid of intron too so watch what
  • 01:05:24
    should be left
  • 01:05:25
    we're gonna expand it a little bit here
  • 01:05:27
    get rid of that one
  • 01:05:29
    exon three so all i did was i
  • 01:05:33
    took and got rid of each exon i mean
  • 01:05:36
    each
  • 01:05:36
    in intron and stitch together
  • 01:05:40
    only the exon so now let's show kind of
  • 01:05:43
    coming out of this process here what am
  • 01:05:45
    i going to have kind of
  • 01:05:46
    popping out off of this the introns
  • 01:05:50
    and when the introns pop off this can be
  • 01:05:53
    intron
  • 01:05:56
    one and then you can have another one
  • 01:05:59
    let's say intron two these are going to
  • 01:06:02
    get popped off
  • 01:06:04
    we don't need these dang things anymore
  • 01:06:06
    so since we don't need them we're just
  • 01:06:08
    going to spit them off during that
  • 01:06:10
    splicing process and
  • 01:06:11
    only lead to the formation of exons now
  • 01:06:14
    within this
  • 01:06:15
    mrna i have my five prime cap
  • 01:06:18
    i have my poly a tail i have only the
  • 01:06:22
    nucleotide sequence which is going to
  • 01:06:23
    code for amino acids
  • 01:06:25
    and then if you really want to go the
  • 01:06:27
    extra mile we said this is the five
  • 01:06:28
    prime in
  • 01:06:29
    this is the three prime end i'm not
  • 01:06:31
    representing any kind of like
  • 01:06:33
    dashes here so this portion here
  • 01:06:36
    and this portion here doesn't get
  • 01:06:39
    translated or red at all by the
  • 01:06:40
    ribosomes
  • 01:06:41
    and so we call these regions since they
  • 01:06:44
    don't get translated
  • 01:06:45
    the five primes it's near the five prime
  • 01:06:47
    untranslated region
  • 01:06:49
    and this one doesn't get translated so
  • 01:06:50
    it's called the three prime
  • 01:06:52
    untranslated region the only portion
  • 01:06:54
    that gets translated is the
  • 01:06:56
    axons now
  • 01:06:59
    i don't know why but they love to ask
  • 01:07:02
    this stuff
  • 01:07:02
    in your exams where you actually go
  • 01:07:05
    through the specific mechanism
  • 01:07:07
    of how the snurps truly do pull the
  • 01:07:10
    n-trons out and splice together the
  • 01:07:12
    axons
  • 01:07:13
    so let's say we take just exon one
  • 01:07:17
    let's write this one as exon one
  • 01:07:21
    and this one is going to be exon two
  • 01:07:25
    and then here in the middle we're going
  • 01:07:27
    to make this
  • 01:07:31
    intron 1.
  • 01:07:34
    so let's kind of show you how these
  • 01:07:36
    snurps
  • 01:07:37
    again the snurps which is the s rna and
  • 01:07:40
    the proteins do this
  • 01:07:41
    so if you really wanted to show it let's
  • 01:07:43
    just represent the snurps as kind of a
  • 01:07:45
    black blob if you will
  • 01:07:46
    they're going to kind of bind
  • 01:07:50
    near this portion here so here's my
  • 01:07:51
    snurp
  • 01:07:54
    within this portion here right at the
  • 01:07:56
    intron at this portion
  • 01:07:58
    let's say here is the three prime end of
  • 01:08:00
    this exon
  • 01:08:01
    five prime end of this exon and let's
  • 01:08:04
    say that here is going to be the five
  • 01:08:05
    prime end of exon two and then the three
  • 01:08:09
    prime end
  • 01:08:10
    of exon two okay and then here's going
  • 01:08:13
    to be the intron inside
  • 01:08:16
    within this intron you have a very
  • 01:08:18
    specific nucleotide sequence that's near
  • 01:08:20
    the
  • 01:08:20
    three prime splice site near exon 1
  • 01:08:24
    and the beginning of intron 1 and then
  • 01:08:26
    you have a very specific nucleotide
  • 01:08:28
    sequence near the 5
  • 01:08:29
    prime splice site at exon 2 and at the
  • 01:08:32
    end of intron 1.
  • 01:08:33
    what are that nucleotide sequence it's
  • 01:08:36
    dumb
  • 01:08:36
    but it helps me to remember it so i say
  • 01:08:39
    i'm a g
  • 01:08:40
    how about you i'm a g so you remember
  • 01:08:43
    g u i'm a g how about you
  • 01:08:47
    i'm a g that's the basic way that i
  • 01:08:50
    remember the nucleotide sequence at the
  • 01:08:52
    three prime splice site
  • 01:08:54
    and then the one at the five prime
  • 01:08:56
    supply site between exon one exon two
  • 01:08:58
    and intron one in this example
  • 01:09:01
    at the there's another one right smack
  • 01:09:03
    dab in the middle let's make him a
  • 01:09:04
    different color so we don't confuse
  • 01:09:06
    it smack dab in the middle there's a
  • 01:09:08
    branch point which is an
  • 01:09:10
    adenine okay there's an adenine right at
  • 01:09:12
    this branch point
  • 01:09:14
    and it has a very specific oh group kind
  • 01:09:16
    of hanging from it
  • 01:09:18
    okay so this is called your branch point
  • 01:09:21
    what happens is is the snurps will come
  • 01:09:24
    in
  • 01:09:26
    and they're gonna cleave at that three
  • 01:09:28
    prime splice site
  • 01:09:30
    okay they're gonna cleave this portion
  • 01:09:31
    off so what would that look like
  • 01:09:32
    afterwards
  • 01:09:34
    so the snurps come in and they cleave at
  • 01:09:36
    that three prime prime splice site
  • 01:09:38
    and so what's going to be left over here
  • 01:09:41
    is we're going to have
  • 01:09:44
    exon 1 somewhat separated here and
  • 01:09:47
    coming off here what's the three prime
  • 01:09:49
    end contain
  • 01:09:50
    an oh group right and again this is exon
  • 01:09:53
    one then the next thing you have here is
  • 01:09:57
    intron one and it's going to have that
  • 01:10:00
    kind of
  • 01:10:01
    portion here kind of split off if you
  • 01:10:02
    will right kind of broken off here
  • 01:10:05
    and then again over here we're going to
  • 01:10:06
    have still fused at this end
  • 01:10:10
    exon 2.
  • 01:10:14
    so again this is my three prime end
  • 01:10:16
    which has that o h
  • 01:10:17
    this is the five prime end of that
  • 01:10:18
    portion of the axon
  • 01:10:20
    and then again same thing over here this
  • 01:10:22
    is the five prime n of x on two
  • 01:10:24
    three prime n of x on two and then again
  • 01:10:27
    what kind of nucleotides do we have
  • 01:10:29
    in here we have that gu which was pretty
  • 01:10:31
    much the marker which the snurp would
  • 01:10:33
    cut at that three prime site
  • 01:10:35
    then on this five prime splice side i
  • 01:10:38
    still have the ag
  • 01:10:40
    and then here in the middle i have that
  • 01:10:41
    branch point with the adenine with the
  • 01:10:43
    oh group
  • 01:10:45
    here's the next thing that happens the
  • 01:10:47
    oh
  • 01:10:48
    group of that branch point will then
  • 01:10:51
    bind or attack that gu site
  • 01:10:56
    and pull it in to where it kind of fuses
  • 01:10:58
    at this point so it makes kind of like a
  • 01:11:00
    little loop if you will
  • 01:11:01
    so let's show that if it attacks the gu
  • 01:11:04
    and pulls it in
  • 01:11:05
    after that happens you kind of form this
  • 01:11:07
    weird little like loopy structure if you
  • 01:11:09
    will
  • 01:11:09
    so what would that look like if we kind
  • 01:11:11
    of droon after we had that attack
  • 01:11:13
    after that attack point it's going to
  • 01:11:15
    kind of look somewhat
  • 01:11:18
    like this if you will where we have now
  • 01:11:22
    that portion where what would be here
  • 01:11:24
    what would be the kind of the nucleotide
  • 01:11:26
    sequence at that point right there g
  • 01:11:30
    and u was attacked at that point
  • 01:11:33
    by the o h at that branch point
  • 01:11:36
    then the next thing happens this is
  • 01:11:40
    crazy
  • 01:11:41
    this three prime o h of exon one
  • 01:11:45
    will then see that five prime splice
  • 01:11:48
    site
  • 01:11:49
    and it'll attack the five prime splice
  • 01:11:53
    site at exon two
  • 01:11:55
    when it attacks it it then breaks away
  • 01:11:59
    the nucleotide sequence ag of this
  • 01:12:01
    intron away from exon two
  • 01:12:04
    so okay now let's show what that would
  • 01:12:06
    look like
  • 01:12:09
    so if the three prime o h attacks the
  • 01:12:12
    five prime n
  • 01:12:13
    three prime o h of x on one attacks the
  • 01:12:15
    five prime n of x on two
  • 01:12:17
    now what do we have here
  • 01:12:20
    exon one fused
  • 01:12:25
    with exon two and we just
  • 01:12:29
    fused the exons and then what do we spit
  • 01:12:32
    out
  • 01:12:32
    after we break this off
  • 01:12:37
    the intron lariat which i showed you
  • 01:12:40
    like that before
  • 01:12:43
    that is how this whole splicing process
  • 01:12:45
    technically occurs
  • 01:12:47
    super quick again snurps bind
  • 01:12:51
    what do they do cut the three prime
  • 01:12:52
    splice side on between exon one
  • 01:12:54
    intron one when it does that the
  • 01:12:57
    o h of the uh adenine at the branch
  • 01:13:00
    point
  • 01:13:00
    attacks the gu site pulls it in creates
  • 01:13:03
    this loop
  • 01:13:04
    the three prime oh of axon one attacks
  • 01:13:07
    the five prime man of axon two
  • 01:13:09
    which snaps the intron out and stitches
  • 01:13:12
    together
  • 01:13:13
    exon one and exon two that is splicing
  • 01:13:16
    you're like zach why the heck do i need
  • 01:13:18
    to know all this crap
  • 01:13:20
    there's a reason why whenever there's
  • 01:13:22
    abnormalities within splicing it can
  • 01:13:24
    produce a various amounts of diseases
  • 01:13:26
    because think about it
  • 01:13:27
    if i don't cut out the introns properly
  • 01:13:31
    and i have introns mixed in with the
  • 01:13:33
    exons and
  • 01:13:34
    introns don't code for amino acids am i
  • 01:13:37
    going to make a proper protein
  • 01:13:39
    no because i'm going to have areas that
  • 01:13:40
    will code free amino acids in areas that
  • 01:13:42
    don't code for amino acids
  • 01:13:44
    you know there's a very devastating
  • 01:13:46
    condition called spinal
  • 01:13:48
    muscular atrophy where they
  • 01:13:51
    are deficient in an smn protein you want
  • 01:13:54
    to know why
  • 01:13:56
    because the snurps aren't working
  • 01:13:58
    properly
  • 01:13:59
    so there's a deficiency or there's a
  • 01:14:01
    problem
  • 01:14:02
    with the snurps not performing the
  • 01:14:05
    proper splicing
  • 01:14:06
    you know what else there's another
  • 01:14:08
    disease called beta thalassemia
  • 01:14:11
    beta thalassemia guess what you don't
  • 01:14:13
    remove a particular intron
  • 01:14:15
    and because you don't remove that intron
  • 01:14:16
    you make a protein that's abnormal
  • 01:14:18
    and it produces beta thalassemia so
  • 01:14:21
    there's
  • 01:14:21
    reasons to know this stuff and again if
  • 01:14:24
    someone has spinal muscular atrophy
  • 01:14:26
    do you know what that affects the
  • 01:14:28
    anterior gray horn neurons and then they
  • 01:14:30
    develop lower motor neuron lesions
  • 01:14:32
    hypotonia hyperreflexia
  • 01:14:34
    floppy baby syndrome right so it's a
  • 01:14:35
    dangerous condition that can be traced
  • 01:14:37
    back to something at the molecular level
  • 01:14:39
    all right now that we talked about this
  • 01:14:41
    there's two more things and i promise
  • 01:14:43
    we're done all right nature so i want to
  • 01:14:44
    talk about two more things and then
  • 01:14:45
    we're done the first thing i want to
  • 01:14:46
    talk about because it's
  • 01:14:47
    very pretty much similar to what we
  • 01:14:49
    talked about over here with splicing
  • 01:14:52
    i want to talk about something called
  • 01:14:54
    alternative
  • 01:14:56
    rna splicing we understand the
  • 01:14:59
    specific reason for splicing it's making
  • 01:15:01
    sure that we
  • 01:15:02
    only utilize exons to code for proteins
  • 01:15:05
    and there's no introns because if we
  • 01:15:06
    have introns in there
  • 01:15:07
    it's going to frack up the whole protein
  • 01:15:09
    production process we'll get an abnormal
  • 01:15:11
    protein
  • 01:15:12
    with alternative rna splicing
  • 01:15:15
    it gives variance
  • 01:15:19
    of a protein and i'll give you guys an
  • 01:15:23
    example
  • 01:15:24
    in just a second but let me kind of talk
  • 01:15:26
    about how this works it's literally the
  • 01:15:27
    same thing
  • 01:15:28
    we're not going to go too ham on this
  • 01:15:31
    let's use the same colors here
  • 01:15:33
    here was exon one and then here we had
  • 01:15:37
    intron 1 and we'll just skip this part
  • 01:15:39
    here where that was intron 2
  • 01:15:41
    and then blue here we had x on 2
  • 01:15:45
    and then at the end here we had in black
  • 01:15:50
    exon 3 okay
  • 01:15:53
    so let's say that we take an example
  • 01:15:55
    here of of of
  • 01:15:57
    this kind of hn rna right so here's our
  • 01:16:00
    h
  • 01:16:02
    and rna and we want to make different
  • 01:16:04
    mrnas
  • 01:16:05
    that'll give variance of proteins so
  • 01:16:08
    let's say here that we have
  • 01:16:09
    i'm just going to put x on one i'm going
  • 01:16:12
    to do all the same color here exon 2
  • 01:16:15
    exon 3 and then here in between we're
  • 01:16:18
    going to have
  • 01:16:20
    intron 1 and intron 2.
  • 01:16:24
    here's what i can do which is really
  • 01:16:26
    interesting and it's very
  • 01:16:28
    cool when it comes to plasma cells and
  • 01:16:30
    antibodies
  • 01:16:32
    so let's say i use those snurps right so
  • 01:16:35
    let's say here i put
  • 01:16:36
    my snurpees right my snurps which are my
  • 01:16:39
    small
  • 01:16:40
    nuclear rival nuclear proteins with the
  • 01:16:41
    snra and the proteins
  • 01:16:43
    they're going to splice but they're
  • 01:16:45
    going to do it in a very interesting way
  • 01:16:47
    so let's say that the first one over
  • 01:16:48
    here we get the same thing that we did
  • 01:16:50
    with that whole process of splicing
  • 01:16:51
    where we got rid of
  • 01:16:53
    all the introns and we only have in
  • 01:16:55
    exons
  • 01:16:56
    and let's say that we have exon one
  • 01:17:01
    let's say that we have exon two
  • 01:17:05
    and then we have exon three right so we
  • 01:17:08
    have all those exons here exon one x on
  • 01:17:10
    two and exon three
  • 01:17:12
    so we'll put these exon three exon two
  • 01:17:16
    exon one so that's one this is going to
  • 01:17:18
    be mrna right
  • 01:17:19
    after we've kind of done that process
  • 01:17:21
    and it'll give way to a particular
  • 01:17:22
    protein
  • 01:17:23
    and we'll call this protein
  • 01:17:27
    a if you will okay then
  • 01:17:31
    we're going to go through the same thing
  • 01:17:32
    the snurps are going to cleave out the
  • 01:17:34
    entrons and only leave in the exons but
  • 01:17:36
    let's say
  • 01:17:37
    for this example we pop out so this one
  • 01:17:40
    we popped out introns
  • 01:17:43
    but let's say with this one we pop out
  • 01:17:45
    both the introns
  • 01:17:46
    and let's say that we pop out x on two
  • 01:17:49
    let's say that we don't want
  • 01:17:50
    x on two in this one so then what am i
  • 01:17:53
    going to be left with
  • 01:17:56
    i'm going to be left with only exon 1
  • 01:18:00
    and exon 3. and by doing that that's
  • 01:18:04
    going to give me
  • 01:18:05
    an mrna that'll code for another protein
  • 01:18:09
    let's call this protein b and then last
  • 01:18:12
    but not least you guys can already
  • 01:18:13
    probably see where i'm going with this
  • 01:18:15
    let's say that this last one example
  • 01:18:17
    three again we cut out the entrance we
  • 01:18:18
    always got to cut out those introns
  • 01:18:20
    but in this case we cut out
  • 01:18:24
    exon three i don't want that one in the
  • 01:18:26
    diagram i don't want this one in that
  • 01:18:27
    mrna
  • 01:18:28
    so what am i left with i'll be left with
  • 01:18:32
    exon one and i'll be left with
  • 01:18:35
    exon two and what will this code for
  • 01:18:39
    this will code for this will give an
  • 01:18:42
    mrna
  • 01:18:44
    that'll then do what code four another
  • 01:18:46
    protein and let's call this protein
  • 01:18:49
    c from one
  • 01:18:52
    h and rna we made three
  • 01:18:55
    different mrnas and made three proteins
  • 01:18:59
    from the same h and rna or from the same
  • 01:19:02
    kind of
  • 01:19:03
    a gene if you will that means it's going
  • 01:19:06
    to be the same
  • 01:19:07
    protein if it's coming from the same
  • 01:19:08
    gene but it's a variant of that protein
  • 01:19:11
    you know what this is examples of think
  • 01:19:13
    about it guys
  • 01:19:15
    think about plasma cells
  • 01:19:18
    which make antibodies when they make
  • 01:19:21
    antibodies you can have antibodies
  • 01:19:24
    that can be secreted or you can have
  • 01:19:27
    antibodies that are different
  • 01:19:28
    and they're expressed on the cell
  • 01:19:29
    membrane that could be one example
  • 01:19:32
    so antibodies differences in antibodies
  • 01:19:35
    would be an example of how that works
  • 01:19:36
    from
  • 01:19:37
    alternative rna splicing because i'm
  • 01:19:38
    making one protein that'll bind to the
  • 01:19:40
    membrane
  • 01:19:40
    and one protein that can be secreted
  • 01:19:42
    think about neurons
  • 01:19:44
    let's say here's one neuron and this
  • 01:19:46
    neuron
  • 01:19:47
    has a dopamine receptor dopamine 1
  • 01:19:50
    receptor
  • 01:19:52
    but then you have another neuron
  • 01:19:56
    and this has a dopamine 2
  • 01:19:59
    receptor it's the same gene that's
  • 01:20:02
    making these proteins but just a variant
  • 01:20:04
    of it
  • 01:20:05
    and then the last thing is take an
  • 01:20:06
    example of a muscle within the heart
  • 01:20:10
    called tropomyosin and the muscle
  • 01:20:13
    and then within the skeletal muscles
  • 01:20:15
    tropomyosin they're different they're
  • 01:20:18
    small
  • 01:20:18
    changes or variants within the protein
  • 01:20:20
    that are coming from the same
  • 01:20:22
    gene so one of the things that they'll
  • 01:20:24
    love to ask on your exam questions is
  • 01:20:25
    alternative rna splicing
  • 01:20:27
    gives you takes one gene one h and rna
  • 01:20:30
    gives you multiple
  • 01:20:31
    mrnas and variants of the same protein
  • 01:20:33
    if you give examples something like
  • 01:20:35
    immunoglobulins dopamine receptors of
  • 01:20:38
    the brain
  • 01:20:39
    or tropomyosin variant within cardiac
  • 01:20:41
    and skeletal muscle
  • 01:20:42
    all right engineers i promise i'm so
  • 01:20:44
    sorry for this being so long but there's
  • 01:20:45
    one last thing that i want us to talk
  • 01:20:47
    about
  • 01:20:48
    the last thing that i want us to discuss
  • 01:20:49
    is called rna editing
  • 01:20:51
    this is also mentioned a lot
  • 01:20:54
    in your exams and the reason why is
  • 01:20:58
    it's it's really interesting kind of how
  • 01:21:00
    this happens
  • 01:21:01
    there's two different types of rna
  • 01:21:03
    editing i only want to mention really
  • 01:21:05
    one of them
  • 01:21:05
    because it's the most relevant to your
  • 01:21:07
    usmles and in kind of a clinical setting
  • 01:21:11
    so let's say here we have our mrna right
  • 01:21:13
    so this is an
  • 01:21:14
    hn rna we've already at this point in
  • 01:21:16
    time for rna editing we've already
  • 01:21:18
    formed
  • 01:21:18
    our functional mrna so at this point in
  • 01:21:21
    time
  • 01:21:22
    this structure here is a mrna
  • 01:21:26
    okay this mrna
  • 01:21:30
    can have a particular nucleotide
  • 01:21:33
    sequence
  • 01:21:34
    that a special enzyme can read and
  • 01:21:37
    sometimes
  • 01:21:38
    switch nucleotides with what is that
  • 01:21:41
    nucleotide sequence which can be seen in
  • 01:21:42
    this
  • 01:21:43
    mrna which we really want to know it's c
  • 01:21:46
    a a we're going to be talking about
  • 01:21:48
    apoproteins that's why i'm mentioning
  • 01:21:50
    caa
  • 01:21:51
    so this is our signal which is really
  • 01:21:54
    really important within this
  • 01:21:56
    mrna which is going to be making april
  • 01:21:58
    proteins a particular protein called
  • 01:22:00
    let's say that this mrna is going to
  • 01:22:01
    code for a particular protein called apo
  • 01:22:06
    b100 if you guys watch our lipoprotein
  • 01:22:09
    metabolism video
  • 01:22:10
    this will sound familiar right but
  • 01:22:13
    april b100 this is going to be the mr
  • 01:22:15
    enable that will code for that protein
  • 01:22:16
    and here's a particular nucleotide
  • 01:22:17
    sequence that we're going to modify
  • 01:22:20
    in the hepatocytes this nucleotide
  • 01:22:22
    sequence
  • 01:22:24
    is not altered in any way it's kept the
  • 01:22:26
    same
  • 01:22:27
    so it's not going to be changed it's
  • 01:22:29
    still going to be c a a
  • 01:22:31
    and whenever this mrna is translated by
  • 01:22:33
    ribosomes
  • 01:22:34
    it makes a particular protein that we
  • 01:22:36
    already talked about called apob
  • 01:22:38
    100 but in enterocytes
  • 01:22:43
    okay your gi cells what are these cells
  • 01:22:46
    here called these are called your
  • 01:22:47
    enterocytes they have a very special
  • 01:22:51
    enzyme
  • 01:22:52
    where they can modify the same gene that
  • 01:22:56
    makes april b100 but make a different
  • 01:22:58
    protein how the heck
  • 01:23:00
    how do they do that let me explain
  • 01:23:03
    there's this cute little blue
  • 01:23:04
    enzyme in the enterocytes called
  • 01:23:07
    cytidine
  • 01:23:11
    d-aminase
  • 01:23:14
    and what the cytidine deaminase does is
  • 01:23:17
    is it deaminates the cytodine right here
  • 01:23:20
    or the
  • 01:23:21
    cytosine nitrogenous base and
  • 01:23:24
    switches it with uracil
  • 01:23:27
    so now let's switch it here where we're
  • 01:23:29
    going to have this as switching c
  • 01:23:32
    and putting u a a
  • 01:23:35
    if you guys know anything about your
  • 01:23:38
    codons
  • 01:23:39
    there's a little trick to remember your
  • 01:23:40
    stop codons you guys remember the
  • 01:23:42
    the little way to remember them you
  • 01:23:44
    remember by
  • 01:23:46
    you go away you are away
  • 01:23:49
    you are gone these are the easy ways to
  • 01:23:52
    remember your stop codons does any of
  • 01:23:54
    these look like a stop codon
  • 01:23:56
    yes uaa ua that's a stop codon
  • 01:24:00
    so what's going to happen is when you
  • 01:24:02
    have the ribosomes which will be reading
  • 01:24:04
    this let's say here i kind of put like a
  • 01:24:05
    little ribosome
  • 01:24:07
    it's going to be reading this and making
  • 01:24:09
    a particular protein
  • 01:24:10
    as it gets to this point where it's
  • 01:24:12
    going to translate it that's a stop
  • 01:24:14
    codon
  • 01:24:15
    will it then read the rest of the rna
  • 01:24:17
    and translate that into a long protein
  • 01:24:19
    no so at this point translation will
  • 01:24:23
    stop
  • 01:24:23
    you won't read all the rest of the mrna
  • 01:24:26
    and make the full protein
  • 01:24:28
    instead you'll make a smaller protein
  • 01:24:32
    and this small protein is called apo
  • 01:24:35
    b-48
  • 01:24:37
    this is something that they love to ask
  • 01:24:40
    on your exams because
  • 01:24:41
    you're taking the same mrna just
  • 01:24:44
    modifying it a little bit
  • 01:24:47
    to produce a different protein that is a
  • 01:24:50
    completely different sized protein
  • 01:24:52
    so that's really cool definitely wanted
  • 01:24:54
    you guys to know that
  • 01:24:55
    and that finishes our lecture on dna
  • 01:24:57
    transcription all right ninja nurse so
  • 01:24:59
    in this video we talk a ton about dna
  • 01:25:01
    transcription i hope it made sense and i
  • 01:25:02
    hope that you guys did enjoy it
  • 01:25:04
    as always ninja nerds until next time
  • 01:25:12
    [Music]
  • 01:25:28
    you
الوسوم
  • DNA-transkription
  • RNA-polymeras
  • promotorregion
  • eukaryoter
  • prokaryoter
  • transkriptionsfaktorer
  • enhancers
  • silencers
  • RNA-splitsning
  • RNA-redigering