title: Early events of RNA polymerase II transcription initiation.
abstract: We have investigated the earliest stages of assembly of an RNA polymerase II transcription complex. General transcription factors from HeLa cells were partially purified and assayed using the adenovirus-2 major late promoter. Preincubation of either all the transcription factors (TF) with the DNA or only the subset consisting of TFIIA, TFIID, and DNA overcame the 15-20 min lag normally observed. The kinetics demonstrate that TFIIA first interacts with the template over a 5 min. period, and then TFIID interacts with the IIA:DNA complex over a 2 min. period. The remainder of the necessary transcription factors then interact with the IIA:IID:DNA complex. There are apparently interactions between IIA and IID, as a pre-incubation of these factors (without DNA) overcomes the lag period. Both IIA:DNA and IIA:DNA:IID interactions are temperature sensitive, resulting in slower kinetics at 0 degree C. Thus, the kinetics of transcription involve activation processes in addition to DNA binding.
authors.affiliation: Biology Department, University of Massachusetts, Boston 02125.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.forename: K
authors.initials: K
authors.lastname: Logan
authors.forename: X B
authors.initials: XB
authors.lastname: Yu
authors.forename: E A
authors.initials: EA
authors.lastname: Davis
authors.forename: M
authors.initials: M
authors.lastname: Posner
authors.forename: S
authors.initials: S
authors.lastname: Ackerman
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D004247:DNA / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000096:biosynthesis / Q000235:genetics*; D013696:Temperature; D013698:Templates, Genetic; D035561:Transcription Factor TFIIA; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000032:analysis / Q000502:physiology; D014158:Transcription, Genetic
chemical_list: D035561:Transcription Factor TFIIA; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D004247:DNA; D012319:RNA Polymerase II