Lecture 14 - Eukaryotic Transcription General Factors and...

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Eukaryotic Transcription General Factors and Mechanism
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Eukaryotes Have Three RNA Polymerases
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Basal Transcription in Eukaryotes B. Transcription Initiation The six general transcription factors for RNA pol II TFIIA, TFIIB, TFIID, TFIIE, TFIIF, TFIIH Plus the Mediator Complex D. Class I and class III factors C. Promoter clearance, elongation, and termination A. Structure of RNA polymerase II
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RNA Polymerase Subunits
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Structure of RNA Polymerase II • Kornberg and colleagues, 2001 • Rpb1, 2, and 3 are homologous to , ’, and of bacterial RNAP • Rpb5, 6, 8, 10, and 12 are found in all three eukaryotic polymerases • 12 subunits in intact RNAP II Names are (R NA P olymerase B ) • Rpb1 includes unique, extended C - terminal domain containing 26-52 repeats of YSPTSPS (important later)
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The Basic Machinery of RNA Polymerase II: Six General Transcription Factors Modified from Callebaut et al. BMC Genomics 2005 6:100
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Basal Promoter Structure in Eukaryotes Smale and Kadonaga, Ann. Rev. Biochem (2003), 72:449-79 • Not all promoters have all elements • Mammals have well -defined start site, but yeast do not
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Model for Assembly of Preinitiation Complex
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Step-by-step Formation of Preinitiation Complex By Class II Factors (1991)
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Footprinting to Define Binding Sites (1990) • Protection appears on both strands from TFIIA and TATA-box binding protein (TBP) from TFIID • TFIIB gives little further change, but one nucleotide (+10) becomes exposed to DNase I • Footprinting by reaction with hydroxyl radicals (from 1,10- phenanthroline-Cu 2+ ) or with DNaseI Experiments used yeast TBP expressed in E. coli
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Model for Assembly of Preinitiation Complex
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This note was uploaded on 04/09/2011 for the course CHEM 4461 taught by Professor Max during the Spring '08 term at Lamar University.

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Lecture 14 - Eukaryotic Transcription General Factors and...

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