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LEC19 - Overall structure very similar to prokaryotic RNA...

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11/12/2008 1 Overall structure very similar to prokaryotic RNA polymerase Assembly of Active Transcription Complexes Prokaryotes: RNA polymerase holoenzyme binds directly to the promoter. Eukaryotes: General T ranscription F actors (TFIID, TFIIA TFIIB) must bind before Pol II TFIIA, TFIIB) must bind before Pol .
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11/12/2008 2 Promoters in eukaryotes contain multiple DNA binding sequences The “core promoter” consists of about 60 bp around the transcription start site Promoters may contain some, all or none of these elements
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11/12/2008 3 l TATA Box: located at about -30 relative to transcription initiation site Is bound specifically by TBP (TATA binding protein) Initiator element: located around +1 bound by specific TFIID Pol II Promoter Sequences (2-3 elements found at most promoters) Downstream promoter element: located around +30 bound by TFIID BRE (TFIIB recognition element) located around -35 bound by TFIIB General Transcription Factors TFIID: TBP TATA bi di t i TBP: TATA binding protein TAF: TBP associated factors TFIIB: stabilization of TBP-DNA interactions, recruitment PolII, start site selection TFIIE: TFIIH recruitment TFIIH: promoter melting, helicase, kinase Mediator: transduces regulatory information from activators and repressors to PolII
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11/12/2008 4 Many sequences and proteins were assayed using run-off assays: in vitro transcription Basal Transcription: Formation of Pre-Initiation Complex
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11/12/2008 5 Unwinding of DNA requires ATP
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