1. Ch 10. Multiple forms of Eukaryotic RNA Polymerase

2. Regulation of Gene expression
Regulatory DNA sequence
Gene regulatory protein

3. Transcription

4. Separation of eukaryotic RNA polymerase
Multiple forms of Eukaryotic RNA Polymerase
Separation of eukaryotic RNA polymerase

5. Cellular localization of the three RNA polymerases
Nucleolus ; pol I - rRNA
Nucleoplasm ; pol II – mRNA
pol III – tRNA, 5S rRNA

6. Roles of Eukaryotic RNA polymerases

7. Sensitivity of RNA polymerases to a-amanitin

8. RNA polymerase subunits (S. cerevisiae)
Complex
2 large subunit + others
5 common subunits a a

9. Human RNA polymerase II
By R. Young, 2000

10. Subunits of yeast RNA polymerase
Immunoprecipitation

11. Yeast RNA Polymerase II subunits
Core subunit : Rpb1, 2, 3 대장균의 RNA pol subunit (b’, b, a) 과 유사
Common subunit : Rpb 5, 6, 8, 10, 12 모든 중합효소에 존재
Nonessential subunit : Rpb 4, 9
Non-classified : Rpb 7, 11

12. Different forms of Rpb1 subunit
RNA pol IIA : bind to promoter
RNA pol IIO (CTD phosphorylation): elongation

13. Structure of RNA polymerase

14. Crystal structure of RNA polymerase

15. Structure of RNA polymerase containing DNA
TATA
TFIIB

16. Promoters Three polymerase recognize different promoters
Regulatory DNA sequence
Gene regulatory protein

17. Class II promoter Core promoter Upstream element BRE
(TFIIB recognition element)
DPE
(downstream promoter
element)
Core promoter

18. Effect of deletions in the SV40 early promoter
TATA box ;
TATAAA(A/T)
-25 upstream from initiation site
TATA-less ; housekeeping gene, - GC box, DPE
Developmentally regulated genes (Homeotic gene, Immune system)
Effect of deletions in the SV40 early promoter

19. TATA box locate the start site of transcription

20. Linker scanning mutagenesis

21. Effects of linker scanning mutations in the promoter

22. Initiator (Inr) ; PyPyANT/APyPy
Downstream promoter element (DPE) in Drosophila ;
30 bp downstream of the Tc initiation site
G(A/T)CG
TATA-less promoter
bind to TFIID
TFIIB recognition elements (BREs) ;
TFIIB for preinitiation complex
(G/C)(G/C)(G/A)CGCC

23. Effects of mutations in GC Boxes on promoter activity
Upstream elements
GC box : Sp1 binding site, orientation-independence, position-dependent
CCAAT (cat) Box ; CTF
Effects of mutations in GC Boxes on promoter activity

24. Class I promoter (rRNA promoter )

25. 5S rRNA promoter (Class III promoter)
Internal promoter : classical class III promoter

26. Promoters of polymerase III genesU6
( Non-classical ; 7SK RNA promoter, a-amanitin에 대한 저항성)
Hybrid (type II + type III ) : 7SL RNA

27. 7SK RNA promoter ; non-classical class III promoter

28. SV40 promoter and enhancer
Enhancer & Silencer
Cis-acting element
Position-independent
Orientation-independent
SV40 promoter and enhancer

29. Effect of deletion in Ig enhancer

30. Enhancer is Position-and Orientation-independent