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Published byKristopher Arthur Hawkins Modified over 9 years ago
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Ch 10. Multiple forms of Eukaryotic RNA Polymerase
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Regulation of Gene expression
Regulatory DNA sequence Gene regulatory protein
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Transcription
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Separation of eukaryotic RNA polymerase
Multiple forms of Eukaryotic RNA Polymerase Separation of eukaryotic RNA polymerase
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Cellular localization of the three RNA polymerases
Nucleolus ; pol I - rRNA Nucleoplasm ; pol II – mRNA pol III – tRNA, 5S rRNA
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Roles of Eukaryotic RNA polymerases
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Sensitivity of RNA polymerases to a-amanitin
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RNA polymerase subunits (S. cerevisiae)
Complex 2 large subunit + others 5 common subunits a a
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Human RNA polymerase II
By R. Young, 2000
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Subunits of yeast RNA polymerase
Immunoprecipitation
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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
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Different forms of Rpb1 subunit
RNA pol IIA : bind to promoter RNA pol IIO (CTD phosphorylation): elongation
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Structure of RNA polymerase
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Crystal structure of RNA polymerase
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Structure of RNA polymerase containing DNA
TATA TFIIB
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Promoters Three polymerase recognize different promoters
Regulatory DNA sequence Gene regulatory protein
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Class II promoter Core promoter Upstream element BRE
(TFIIB recognition element) DPE (downstream promoter element) Core promoter
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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
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TATA box locate the start site of transcription
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Linker scanning mutagenesis
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Effects of linker scanning mutations in the promoter
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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
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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
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Class I promoter (rRNA promoter )
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5S rRNA promoter (Class III promoter)
Internal promoter : classical class III promoter
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Promoters of polymerase III genes
U6 ( Non-classical ; 7SK RNA promoter, a-amanitin에 대한 저항성) Hybrid (type II + type III ) : 7SL RNA
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7SK RNA promoter ; non-classical class III promoter
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SV40 promoter and enhancer
Enhancer & Silencer Cis-acting element Position-independent Orientation-independent SV40 promoter and enhancer
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Effect of deletion in Ig enhancer
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Enhancer is Position-and Orientation-independent
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