biol321 molecular genetics - wordpress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 upstream...
TRANSCRIPT
![Page 1: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/1.jpg)
1
Biol321 Molecular Genetics
Handout 9
![Page 2: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/2.jpg)
2
Yeast RNA polymerase II subunits
PTSPSYS CTD
Epitope tagging
![Page 3: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/3.jpg)
3
Prokaryotic RNA polymerase
Eukaryotic RNA polymerase
RNAP from T. aquaticus
![Page 4: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/4.jpg)
4
Initiation of transcription by RNA
polymerase II
UCE
-200 -150 -100 -50 +1 +50 +100 +150 +200
-45 +20
-25 InrUps tream Elements
Core promoter
RNA Pol I
RNA Pol II
RNA Pol III
RNA polymerase promoters
![Page 5: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/5.jpg)
5
Initiation at RNA Pol. II promoter
Initiator sequenceYYCARR
-25 box = TATA boxTATAWAW
-25 InrUps tream Elements
RNA Pol II
TFIID: (~800kD, larger than RNA pol II at ~600kD)
TBP
TAFs(9 to14)
TBP = TATA binding protein
TAF = TBP associated factors
TATAAATATAAA
mRNA
Building the preinitiation complex (PIC)
![Page 6: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/6.jpg)
6
TBP bends DNA ~80o and forces open the minor groove
![Page 7: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/7.jpg)
7
-200 -150 -100 -50 +1 +50 +100 +150 +200
-25 InrUps tream elements
RNA Pol II
Initiation at RNA Pol. II promoter
TF TF TF
- + +
TAF
TBPA B
F
EH
Stimulation of transcription by activator binding to an enhancer
![Page 8: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/8.jpg)
8
The Mediator Complex Links Activation Domains to RNA Pol II
Activation of transcriptionBy transcription factors (TFs),
activator, and coactivator proteins
![Page 9: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/9.jpg)
9
Initiation at RNA Pol. I promoter
Core Promoter
Upstream Control Element
(CP)
Initiation at RNA Pol. I promoter
(CP)
RNAP I
UB
FU
BF
UB
FU
BF
SL1TIF-IATIF-IC
OnUBF UBF Binds (as a dimer) CP and UCE
SL1 SL1 (in humans) ≡ TIF-IB in mouseTIF-IATIF-IC Recruitment of RNA Pol. I
RNAP I RNA Pol. I
![Page 10: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/10.jpg)
10
RNA polymerase III promoters
Initiation at RNA Pol. III promoter
TFIIIC Binds box A and box C
Binds to TFIIIC
Recruitment of RNA Pol. III
RNAP III RNA Pol. III
TFIIIA
TFIIIB
Recruit TFIIIB
TFIIIC TFIIICTFIIIA
TFIIIB
RNAP III
On
![Page 11: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/11.jpg)
11
![Page 12: Biol321 Molecular Genetics - WordPress.com · 7-200 -150 -100 -50 +1 +50 +100 +150 +200 Upstream elements -25 Inr RNA Pol II Initiation at RNA Pol. II promoter TF TF TF-+ + TAF A](https://reader034.vdocuments.us/reader034/viewer/2022052500/604932a1981d1167c161e1d1/html5/thumbnails/12.jpg)
12
Regulatory
Holoenzymes with different promoter-binding specificity
SigmaFactor Promoters Recognized Promoter Consensus
-35 Region -10 Region70 (rpoD) Most genes TTGACAT TATAAT32 (rpoH) Genes induced by heat shock TCTCNCCCTTGAA CCCCATNTA
-24 Region -12 Region54 (rpoN) Genes for nitrogen metabolism and CTGGNA TTGCAother functions