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Lecture03 01 P24 RNA Synthesis

Apr 06, 2018

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    Lecture 03/ 01P24

    RNA SYNTHESI S

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    -

    Just one of t he DNA st rands for a gene

    called the templatestrand.

    molecule need not use t he same st rand

    as the tem late. RNA polymerase is t he enzyme that

    uses DNA as a t em late t o make RNA.

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    I nit iat ion of t ranscript ion requires t hatRNA ol merase reco nize and bind

    t ight ly, apromotersequence on DNA.

    Bindin can be inf luenced b other

    proteins. The romoter se uence direct s t he RNA

    polymerase as t o w hich of t he t w o

    st rands is t he template and t herefore inw hat direct ion t he RNA polymeraseshould move.

    See Figures Figure 14.7 Part 2 and 14.4 Part 1

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    .

    Part 1

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    .

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    .

    Part 2

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    ,t he DNA about 10 base pairs at a t ime

    init iat ion sit e) in t he 3 - to-5 direction.

    t he 5 to 3 direct ion; i.e. t he RNAa sc p s a pa a e o etemplat e st rand.

    See Figure 14.4 Part B

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    .

    Part 3

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    ,aw ay, allow ing the already t ranscr ibed

    helix.

    e energy or syn es s comes rom

    the removal and breakdow n of t hepyrop osp ate group rom eacnucleoside t r iphosphate building block

    added. (See Figure 11.21 Part a, Edn 7)

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    Figure 11.21 Sequencing DNA

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    art icular base se uences

    ar cu ar ase sequences n eDNA specify t erminat ion.

    I n eukaryotes, t he process

    does not occur unt il a sequence

    polyadenylat ion has been

    transcribed (See Figure 14.4 - Part C)

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    .

    Part 3

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    There are t hree major classes of RNA: essenger m - co es or a

    protein

    Ribosomal RNA ( rRNA) - cont ribut es t ost ructure and funct ion of r ibosomes(protein synt hesis machinery)

    Transfer RNA t RNA carr an aminoacid and act as an adapt or moleculeallow in decodin of mRNA to rotein

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    Prokaryot es have one type of RNA,

    t RNA, and rRNA

    polymerases: I , I I , and I I I .

    eukaryotes.

    -RNA polymerases occur for every 104

    t o 105 bases incor orated.

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    DNA codes for RNA by t ranscr ipt ion.

    nucleot ides (codons) . Since t here are

    possible codons.

    .

    The 64 possible codons code for only

    signals found in all mRNA molecules.

    See Figure 14.6

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    .

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    One mRNA codon, AUG ( t he start

    codon) , indicates t he start ing point oft ranslat ion and codes for methionine.

    and AUG must t herefore be used w hen amet hionine occurs w it hin a prot ein.

    Three stopcodons (UAA, UAG, and

    reading the mRNA; t hat is, t hey.

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    After subt ract ing start and stopco ons, t e rema n ng co onscode for 19 di f ferent amino acids.

    This means t hat many amino.

    Thus t he code is redundant.

    . .

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    The code is not ambiguous. Each codon,t w o or t hree possible amino acids.

    e genet c co e s near y un versa ,applying t o all species on our planet .

    Minor variat ions are found w it hin

    mit ochondr ia and chloroplast s; otherexcept ions are few and slight .

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    u aryot c un e pro aryot cDNA) , is separated f rom the

    cytoplasm by being contained w it hina nucleus.

    The init ial mRNA t ranscr ipt of t he

    DNA is modif ied rocessed beforeit is export ed to t he cytoplasm.

    ev ew gure . art

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    .

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    After t ranscr ipt ion, t he pre-mRNA is

    -G cap at t he 5 end and a poly A t ail

    t he AAUAAA sequence).

    t he stabilit y of t he RNA and for it s

    Review Fi ure 14.10

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    .-

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    The int rons are removed from t he mRNA,of RNAs and proteins.

    s soon as t e pre-m s t ranscr eseveral small nuclear r ibonucleoprot einpart c es sn s n

    This links all t he coding regions t ogetheras a cont inuous sequence.

    .

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    .

    Machine

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    be cont rolled at t he:

    ,

    posttranscriptional,

    t ranslat ional, and

    .

    .

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    .

    Ex ression Part 1

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    .

    Ex ression Part 2

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    .

    Ex ression Part 3

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    e ma or me o o con ro s se ec ve

    t ranscript ion, w hich result s from specif ic prot eins

    t ranscri t ion factors bindin t o re ulat orregions on DNA.

    Specific t ranscript ion fact ors must bind t o t hepromoter e ore po ymerase can n .

    A group of general t ranscript ion factors

    w ork w it h RNA polymerase I I t o t ranscribe

    mRNAs I nit iat ion also depends on t he proteins bound(act ivators and repressors) .

    Review Figures 16.14 and 16.15

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    .

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    ., ,

    Activators

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    The simult aneous cont rol ofw e y separate genes spossible t hrough proteins t hat

    bind t o common sequences in

    Review Figure 16.17

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    .

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    -

    Some 50% of eukaryot ic genesgenera y ave severa exons analternativesplicing can be used t o

    produce dif ferent prot eins.

    Review Figure 16.22

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    .

    Mature mRNAs and Proteins

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    -

    The stabili t y of mRNA in t hecytop asm can e regu ate ythe binding of proteins.

    Specif ic AU-r ich sequences mark

    by a ribonuclease complex ( t heexosome

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    ene expression

    One class are microRNAs miRNAs- short , 19-25 nucleot ide, non-codin RNAs

    Some miRNAs bind complement ary

    and prevent t heir t ranslat ion and

    .

    ev ew gure .

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    .

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    ene ex ression

    t he human genome is st i l l unknow n;current est imat es ran e from 500

    1000 -

    genes; t hus on average each miRNA

    might regulate about 200 t arget genes Many genes have target sit es for a few

    dif ferent miRNAs

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    miRNAs appear t o be de-regulated in in several

    including chronic lymphocytic leukemia, pediat ric

    Burki t t s lymphoma, gast ric cancer and lung cancer

    The miR-17-92 miRNA clust er on chromosome 13is fr equent ly ampl if ied in B-cell lym phomas

    Elevated levels of t hese miRNAs are found inclinical samples and in related cell lines

    When t hese miRNAs are overexpressed in amouse model cancer resul t s

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    Conf irm that many miRNAs are t issue-

    s ecif icall ex ressed Potent ial for diagnost ic use:

    -

    - dist inguishing bet w een relat ed cancers

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    Oft en t here is lit t le correlat ion bet w eene a ou o a e a ou o

    protein synthesized.

    be determined by factors act ing aft er

    . One w ay is binding of repressor proteins t o

    .

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    -

    Regulat ion of protein longevit y:

    modified aft er t ranslat ion.

    w ay to cont rol it s act ions.

    l inked to a 76-amino acid prot ein,

    . Other ubiquit in chains t hen at t ach t o the

    .

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    The prot einubiquit in complex t hen binds

    .

    I nside the proteasome, energy f rom ATPs use o cu o e u qu o

    recycling) and unfold it s t argeted protein.

    Proteases digest t he protein int o smallpept ides and amino acids.

    .

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    .