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Note10 - Lecture 11 S Harmer Basal Transcription machinery...

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Lecture 11 MCB 161 S. Harmer 02/09/2010 Basal Transcription machinery Reading: Watson: 394 - 412; 415 - 416; 457 - 458; 777 - 778 Problems: see web site. I. Component identification for Pol II transcription A. Mutagenesis identifies promoter elements B. Biochemical fractionation of transcription factors II. The preinitiation complex and DNA interactions A. TFIID 1) Footprinting to define binding site 2) Gel shift assays to detect DNA binding 3) TBP, DNA binding, and conformational change 4) Functions of TAF II s B. Sequential formation of the preinitiation complex (PIC) 1) TFIID 2) TFIIB (TFIIA), gel shifts to evaluate binding 3) Pol II, TFIIF 4) TFIIE, TFIIH C. Promoter clearing and elongation 1) Promoter melting 2) Pol II C-terminal domain phosphorylation From: Nikolov, D. B. and Burley, S. K. (1997) RNA polymerase II transcription initiation: A structural view. Proc. Natl. Acad. Sci. USA 94 :15-22. Note: TFIIA subunits are included among "TAF II s" on this list.
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Lecture 11 MCB 161 S. Harmer 02/09/2010 Example applications of gel shift and footprinting in study of basal apparatus assembly.
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