Cox J M, Hayward M M, Sanchez J F, Gegnas L D, van der Zee S, Dennis J H, Sigler P B, Schepartz A
Department of Chemistry, Yale University, New Haven, CT 06520, USA.
Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13475-80. doi: 10.1073/pnas.94.25.13475.
By selective attachment of a DNA cleavage agent to specific residues in the yeast TATA box binding protein (yTBP), we demonstrate that, in solution, yTBP binds to the TATA boxes of both the adenovirus major late promoter and the yeast CYC1 promoter with only a modest preference in orientation and binds well to several overlapping binding sites. The general factors TFIIA and TFIIB each increase the rotational and translational selectivity of yTBP but are not sufficient, at least individually, to confer a unique polarity to the preinitiation complex. We conclude that TBP alone cannot define the productive orientation of general factor assembly on a promoter.
通过将一种DNA切割剂选择性地附着于酵母TATA框结合蛋白(yTBP)中的特定残基上,我们证明,在溶液中,yTBP与腺病毒主要晚期启动子和酵母CYC1启动子的TATA框结合时,在方向上仅有适度偏好,并且能很好地结合几个重叠的结合位点。通用因子TFIIA和TFIIB各自都增加了yTBP的旋转和平移选择性,但至少单独来看,它们不足以赋予起始前复合物独特的极性。我们得出结论,仅TBP本身无法确定通用因子在启动子上组装的有效方向。