Topological localization of the human transcription factors IIA, IIB, TATA box-binding protein, and RNA polymerase II-associated protein 30 on a class II promoter.
The human general transcription factors IIA and IIB bind directly to the TATA box-binding protein (TBP), and modulate transcription initiation by RNA polymerase II. RAP30, the small subunit of TFIIF, binds to TFIIB and RNA polymerase II and recruits RNA polymerase II to a preinitiation complex containing TBP and TFIIB. By using the adenovirus 2 major late promoter tagged site-specifically with the photoactivatible cross-linking reagent N3R-dUMP we have localized TBP, two subunits of TFIIA (A35 and A21), TFIIB, and RAP30 along promoter DNA. TFIIA cross-linked to the coding strand opposite TBP at the TATA box and cross-linked upstream of TBP around position -40. RAP30 cross-linked strongly and TFIIB weakly to the coding strand just downstream of TBP at -19. We interpret these data in the context of a molecular structure for the TBP promoter complex.
Coulombe
B
B
Banting and Best Department of Medical Research, Toronto, Ontario, Canada.
Li
J
J
Greenblatt
J
J
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003432:Cross-Linking Reagents; D018360:Crystallography, X-Ray; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010777:Photochemistry; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D012319:RNA Polymerase II / Q000378:metabolism; D016385:TATA Box; D035181:TATA-Box Binding Protein; D035561:Transcription Factor TFIIA; D035581:Transcription Factor TFIIB; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D019473:Transcription Factors, TFII
D003432:Cross-Linking Reagents; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D035181:TATA-Box Binding Protein; D035561:Transcription Factor TFIIA; D035581:Transcription Factor TFIIB; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D012319:RNA Polymerase II; C056604:transcription factor TFIIF