title: Dynamic architecture of a minimal RNA polymerase II open promoter complex.
abstract: The open promoter complex (OC) is a central intermediate during transcription initiation that contains a DNA bubble. Here, we employ single-molecule Förster resonance energy transfer experiments and Nano-Positioning System analysis to determine the three-dimensional architecture of a minimal OC consisting of promoter DNA, including a TATA box and an 11-nucleotide mismatched region around the transcription start site, TATA box-binding protein (TBP), RNA polymerase (Pol) II, and general transcription factor (TF)IIB and TFIIF. In this minimal OC, TATA-DNA and TBP reside above the Pol II cleft between clamp and protrusion domains. Downstream DNA is dynamically loaded into and unloaded from the Pol II cleft at a timescale of seconds. The TFIIB core domain is displaced from the Pol II wall, where it is located in the closed promoter complex. These results reveal large overall structural changes during the initiation-elongation transition, which are apparently accommodated by the intrinsic flexibility of TFIIB.
authors.affiliation: Department of Chemistry and Center for Integrated Protein Science München, Ludwig-Maximilians-Universität München, Butenandtstr.11, 81377 Munich, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Treutlein
authors.forename: Adam
authors.initials: A
authors.lastname: Muschielok
authors.forename: Joanna
authors.initials: J
authors.lastname: Andrecka
authors.forename: Anass
authors.initials: A
authors.lastname: Jawhari
authors.forename: Claudia
authors.initials: C
authors.lastname: Buchen
authors.forename: Dirk
authors.initials: D
authors.lastname: Kostrewa
authors.forename: Friederike
authors.initials: F
authors.lastname: Hög
authors.forename: Patrick
authors.initials: P
authors.lastname: Cramer
authors.forename: Jens
authors.initials: J
authors.lastname: Michaelis
mesh_terms: D031541:Fluorescence Resonance Energy Transfer; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D012319:RNA Polymerase II