title: Formation and fate of a complete 31-protein RNA polymerase II transcription preinitiation complex.
abstract: Whereas individual RNA polymerase II (pol II)-general transcription factor (GTF) complexes are unstable, an assembly of pol II with six GTFs and promoter DNA could be isolated in abundant homogeneous form. The resulting complete pol II transcription preinitiation complex (PIC) contained equimolar amounts of all 31 protein components. An intermediate in assembly, consisting of four GTFs and promoter DNA, could be isolated and supplemented with the remaining components for formation of the PIC. Nuclease digestion and psoralen cross-linking mapped the PIC between positions -70 and -9, centered on the TATA box. Addition of ATP to the PIC resulted in quantitative conversion to an open complex, which retained all 31 proteins, contrary to expectation from previous studies. Addition of the remaining NTPs resulted in run-off transcription, with an efficiency that was promoter-dependent and was as great as 17.5% with the promoters tested.
authors.affiliation: Department of Structural Biology, Stanford University, Stanford, California 94305, USA.
authors.forename: Kenji
authors.initials: K
authors.lastname: Murakami
authors.forename: Guillermo
authors.initials: G
authors.lastname: Calero
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Brown
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.forename: Ralph E
authors.initials: RE
authors.lastname: Davis
authors.forename: Hinrich
authors.initials: H
authors.lastname: Boeger
authors.forename: Roger D
authors.initials: RD
authors.lastname: Kornberg
mesh_terms: D004271:DNA, Fungal / Q000737:chemistry* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000502:physiology*; D012319:RNA Polymerase II / Q000737:chemistry* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: D004271:DNA, Fungal; D014157:Transcription Factors; D012319:RNA Polymerase II