Lecture 14 Notes
Lecture 14 Notes BIOL-L211 2521
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This 14 page Class Notes was uploaded by AnnaClippinger on Wednesday October 21, 2015. The Class Notes belongs to BIOL-L211 2521 at Indiana University taught by Megan Dunn in Summer 2015. Since its upload, it has received 36 views. For similar materials see Molecular Biology in Biology at Indiana University.
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Date Created: 10/21/15
Lecture 14 39 Oct 15 2015 39 Announcements 5311 Honors section of Genetics is offered this spring 39 LM in GPA of33 39 39 Last Time Transcriptional Regulation in Prokaryotes Examples of Prokaryotic Gene Expression End of Exam 2 Material Today V l1iquotl quot A n i l39 Ii 139 I I 7 IL 39f39 z Leial quot55 9F Start of Exam 3 Material l i l39 in Transcription in Eukaryots 391 Next Time Optional Review Session on Tuesday Exam on Wednesday from 79 PM in BH 013 Bring pencil and student ID m w t e s w to stir inn Tronscn non in eukor ores evl39l ml OHVPA s l Ual lqllll all 24 m p V q dig12mm snafu 4 lil r litl wt o iie o ation nd termination quot Upmfm gl xw mjhl will UN in mum seal M Mien noquot l Miriam genie5 s Im Pads than le39llHNtf Transcriptran accursrn 114ch mg Mumnae It quot tmn lntwn lg lg C l DPl GM V aquot than 39 sinquot mm Us 1 l Alt quot1 7i m a y a l g a l 1 1 n i i mi F Fr it i l Hf a aquot 7 xx PmcessingJi 3 7 r Eli MVM IHNE 9 Mainstream I i a mid transpdnl Mlll39 um I quotarmling V v 539quot polypeptide 7 growing polypeptide Fro kanrotes Eukawotes Transcrip orr in eukaryotes Mr M WW i3 primarily used IQ 35 l quotl0 Region u Multiple general tra navigation factors involved ii j i L 39m PPM qpll39cmg 10 mom Iquot 3 PNldm M O a V ll39n I gs I ml39eI Beg rudn cn I mlhalN 1 htlpz bmbvpsuedufdirector jer jl T23quot D D Em llt am Ml c Jami haw Mfr o g 5 N0 Structure of a eukaryo c gene Recall that eukaryetic39genes consist Dfexons and introns r l I MBOTH are pmzm in im Prll39l39lllW 2239 l lran i CVIPh cmg 39 mq naming I Separate coding n Will be both transcribed into mRNA and h quot 39 i 39 then translated into protein Md lbw I m xi ifii39 if I l 7 gt quot V A I w l TYMGCI I39Pl39lon RNA x IranScript spliced mRNA 5 13 Fig 843 Transcriptional control of a eukaryotic gene we t 9 l i L 1 EL l l 0 n c39NP on fact0Y9 WWHre m 5 m 39 twang 1393quotqu red I if help QM Heb RNA P m5 9w l a Vary depending on location in n cti on organism Bind regorlaor oroteins like activators anal repressors r can lot u gt m or downstrcgm of C l l 96 WM Hund quotd9 0F News 9 an Fram can Enhancer Proximal Falwell metal control central Transcription signal TENEF N FI M elements elements etart site sequence t Erml Et lm Inn 3 Emu Vlntren Enon intronEsen gram 7 5 n r 3quot quot 39quot if if 397 if WV 739 397 i U stream 39 m 39r 3 l P lT f FDIY ADM nstrea39n I signal Primary are E Intreer Error jlntrgn Em L 7 Cleaned traneerift quot1 Tender pram im srimary ranserim PWET lilil proeeesin 7 7 lntron Hil g a T39 r i a l Endings femur r r ligr 39 003 mRN e revere 39m39 l b 39 Hi 3quot Ll rE I 39 imm5mf rr l E umr r n e pmm onr n WesUT R39W Ia if i N 039 I MEMMMM Similar F 0 1 o e an target a C a 55 a as SIMplo as Pratt 3W W6 00 Frommquot Structure of core promoterquot a ll c l l an A setoi seouence sl eci c elements needed by RNAP to initiate transcription Elements colored in xeslcan be upstream or downstream of the t t ra n at ri pti o nal sta nit site i WGHQCHP39I l w l g 3 l FI ETBEE TFiIDi IEHQ TFlgj uTFllD slr a2 1 26 16 21 2na 30 32 34 ll l TEFE TATE l quot i if I H i l an G G l l Tn been Tet ETTC been ii a a l quotAGE CCACGLCC anlg q T Tm TT GGTCGTG V l T i 39wIHRFiiii isg c 39CUWQ39CHGUG wwuenceq all the 1 once LO W a quot Promoters include some combination but not all of the elements shown 39l The nag RNA poiymemsw wmtw Mm WWW Lg fucusmg MI All eukaryot CHM 5 es haire at least 3 RNA polymerases lisTranscm ihe SpecializedRNA encoding genes gt marm 39meat n quotEh39wdd quot 39 w illfocuson Phl l39l39l 39 V quot39 39 Plants also have Pol IV and Pol V 47315jhfs made up of mu tiple subunits V WWI of RNA F6 FcQLMMOS crab GMW Transcription initiaiign involves three teps Ending closed xmmpmx prmnoler39 quotmeltingquot I if 39 Men unmet 1 i niliat transcri ing cwplex f quot dmmsmraam gig luatmn ILW quotf A a L r9 0 fi mt IO N UC60rde9 x The preini e on complex forms at the promoter 21 ajtieajag Pol II 6 general transcriptien factors TF5 add itionai proteins bound at the promote V r Ht 1P9 FWH Wquot PM E ever were crlp umt G mrt arte Gammacc DNA quot Grands 9 Pmmntw i i Recruitment to premiereth cempieXpmmeter Mullthubum t 7 ranscriptien factor for Pol II D CUM Matti 5n bunit protein contains th tnisij39xtgggt it V Z WK 1 we we TATP From 0 RF Tee TATE hex TFIMQ 39 Mark mum 4 transcription fewer Rec mn rt element int Ha roe 6 arm n claimant Fig 13 15 stabilizes roteinpmtein interactions i xiiir which at 14m f ti39l Wm 91 lnvolvcam gment 10 tuntort have I Silni afunch39on g 7 The premr atmn complex forms at the promoter T N 05le 45 I I36 I TH mi Me 039 39t 5 Sim tartan Pam marge Pol Ii 5 general transcription factors adciitional proteins bound t titan Prion tumors Heips position Pol il over transcriptional start site separates DNA strands at promoter I r Gtarrs downstm m Joins the complex recruits TFIiH quot i M trimtut Promo r if Hard relives r Fig 1316 10 Additional components of tire preim39tt ott39on complex I 7 M11 mate 1 re that L quotV 0 39ind enhahcers upstream of promoter helprecr uit Pol39ll t5 promoter I r r 0le amvaro Remember Eukaryotic DNA is compacted into mucleosomes v V H M Pm WNW HM COMPan Must gum access 11 DNA 1T0 HGorrptron n prorch Modrw or remodel NHGI MQUMQEQ Note that ma my pictures in chapter don t Show Mediator complex in the cartoons Fig 1319 11 t Aborgtiveinitiationf nd promoter escape 39 Involves two stepg TFIIE M nd dawmtwqm OF pol Holdq Fromm rpm l I 93965 MIR 1 drawer to 440 mm mm a trandcts cPamm Before the second step in promoter escape occurs I r 7 Pom eatarr t tha promom NMWIEGS Numerous Gumr tranea rme a Made 3 dr Upped Made 1 drop Abvort ive initiation and promoter escape Nic i 1 39 I c am plum Helm P0111 e cdmquotm enmul mnscHNrm 4 can MW b tanairs Fig 1316 n 13 Elongation K No longer need museq Initiation factors and Mediator complex Esa aced H 1 0W 90 d ow namely 2 E i am am 7 f the mRNA transcript can begin d A The same hasi base pairing appliieezi RNA nucleotides base pair with DNA x l N T 6155 ei umgatien ii quot Elongation The same chemical reaction of transcription occurs as with prokaryotes B and v phosphates removed rorn incoming r39iioonucleotidie 7 i i PMUGPha dic Er e 39 Fumed anOMN ade 2 Manama NTF 14 V E ukaryoo39c chromosomes are compacted by histones Ma kc Hquot More com Pli care0i DNA must be compacted to t in the nucleus W DNA i W uhd it Nucleosm This is mediated by proteins calle TEIOII39IIEI E nuclear Fare Condensed Chromosome i 39 complex of and protein h r A single unit of DNA wrapped around an octamer of histories 397 39 V However this can 39oseaotentialissuei 7 N c DUrm Eliaquot paHmEi e39w recommwm39wcm WM r 4 The nucleosome How does transcription proceed past nucieosomes J I a li quot n r First FEWEW what nucleosomes are 391 4 singie D NA hlstone octamer core complex H3 H1 290 WELL 1 H3 and form a etrr1rnerr HZA and HEB form a dimer 4 Lap er 4 copra 2 H3H4 tetramer binds dsDNA 39 39 FCVWQ as ho1 Mi l l l Amt1w wrap the DNA 339 Two HZAHZB dimers join the complex Ijh e nucleosome is formed Fig 320 15 quot quot neg cnmrri c the Defamer HAH5 FACT aids elongation in the presence of chromatin v H mdlmw OF 1 i Binds H 39 H 13 I maquot V IlEAH2 B dimer 7 L which allow9 m p111 W 391 Warm step 1 M hisionesrl H EA I H28 H3 H u HZA39H2B dimer 513p 2 N o 39 aw cloned Pm all disemscmwd 971m 9 mm as uowswo mc c a Fig132 3 s ii hlelpe in nucleooome reassembly 17 RNA processing factors faET DYG mm m Wde in e Wm i WWW WWW 41m m n Splicing comipllicatedwe will spend separate Iectures on this a Polyadenyla on of3 end 39rocess mRNA 7 7 quot In M qgsmwm 397 o no Mk MRNA ids gc spiking machinery b phosphoma on stage a FINA processing 51am of cm tail transcription tasters cm39 lzad 2 3 4 gt a g MPFMQ ocean as Goon am 1 quot 39l rangicw p1 emerges from F0111 E i quot quotQ E c mammal mm 43 on Iquot PT 34 P TT 3 a was Figi 13 21 1398 Formation of the 5 cap a 394 M Wl m 9mm E9 add ma G39cnd a RN45 i 39 Jg glnw ver hreequot gte 5 Helpewmarw Mk I munm Q e M 0amp RN H G 39 F 19 Fofma on of the E mp I I A 39 n thvlated guanine is added m the 5 end of RNA 39 Ivolafes three steps 20 Forma on of the 5quot crop A methylated guanine is added to the 5 end of RNA Involves three steps Equot m Pa Eminch OF RNA from degrgd 9qu Also helps with translation initiation 39 39 LlMMge f r WNIWE 10 Mcmviaiul guanine in RNA CTD toil recru s polyodenylo on enzymes Mama W 096 Wl twmmaI Hm fltl39is if 4L l ii l L s ll quot i l aquot 39 HRH 5 anyquot at W if tzr F5 gigai at a F but I 5quotan dollsa in jug lil I ll l l I1llll t lfiu39 La lulu n M 1 M 7 Mv v capping enzyme polyadenylation 39 and cleavage quot factors components of splicing machinery As 39 transcribes toward theof a gene 1 1 ssociated with polivadenvlation enzyme5K quot quot transcribed not 5 own in this guresee next slide ad en 1 may mt W mm TD 391ka in l irequotEU in yll u r vf l l 22 412 mu Tran i Lead In Edy m mama As Pol II transcribes toward the end of a gene Polyadenyla on of 3quot end V w h 7 397 7 7 r F 5 Jquot T KILE E ii rh 39 quot I g r 1 3 I 39 39 F 1 1 cleavage stimltion fatste 39quot cleavage and polyadenvlatiorm speci d y factor 1 Whm mnqcrl39kd m mes 69 l 0 F91 hm nefcr From CTDTAI I 1 D mph puttyA signal sequencemDNA 54 7 polyaAsi39gnal 39 sequencein RNA 5mm v RNA 23 Polyadenyla on of 3quot end punyAEignal sequa ce MEDINA ids H mRNA cl ca we my RN A In 10 1 Frag Ghor39mmd will be dearanJ Dissociates I pawA sigma V Hsequence in RNA E39GA n r a RNA RNA cleavage I 39 P SF E GAP polwlxpal39ymems 39 r PAP 5 CAP nde39l39D m RNA 9 PM ah rgml Rnc Bird s 0 3 Em PAP 39 7 3 f 241 Polyadenylation 01 3quot end pm l KdaIwn pmwmmg i linds 3 end of mRNA l v r J Adda a rm alEMm m a39 Nu lw d h the and U c mn9crtyiLj z39 9 nlnlyrAfbinding prniein 1 SOAP 7 additional polyAn binding pvmtein he nclPGPmM l Bind t0 and stabilize pelyv Atail involved In M RNA mam ii mm la nn tailquot J39i39 iquotE39Ei quotGiy39 6 Ii rkii39s jEpic J Ll VJ i 3 l 39 he ll lu 539 CAP 39 Hg 13 25 25 Transcriptional termination Torpedo model Fa rc 6 T I w it A r A v i in 5 a 511 71 an 1 I rf quotiquotll mil 3 r 13 it 1 l relLL I gillEffi39injJ1 lgquot J L143 quot 4thquot 397 39g an may 1quot z39 3quot quotr quottquot fi239f I H MIL 34 than Note polyA git is light green i UAsevuence an in transcript pallyA signal x39 saquenceinEDNA 39 a 2211quot f39iquotquotllfrifr I a mRNA is cleaved into two pieces by CstF 39 The smaller piece is degraded by an 5 7 ll iiiquot RNA polyA Big nai sequence in RNA The torpedo modequot Suggests RNA cinavaqa 30 TM6 KNML fhatdegradcs 1 v 5m er Pica Maxi wrggw matmn m r g f W WW 7 ttltl pmi39mmii WM man an 5 CAP 2395 Transcriptional termination Torpedo model Note poly A site is light green in DNA sequence and in tra nscriptl Fellow and understand each picture step by step p lyA signal Rat lernz sequence f a Cleavage by CstF NA Ma 393 4 aquot dirccrtm until it mam Pol Pal II d39Iqsmrawr 27 Transcriptional termination Ailosteric model Note polyA signal is light green in DNA sequence and inlltlranscriptl mgmg tamquot mm gmquot L r quotn 11 clmvagc f 39 PM ECMHSE tunfurmah39on dccreaqc In yucca iwi e l GPOWMWM v dil socm w 28
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