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A comparison of the DNA bending properties of the DNA binding proteins CRP and TFIID

Gaston, Kevin; Bell, Andrew; Busby, Stephen; Fried, Mike

Authors

Kevin Gaston

Andrew Bell

Stephen Busby

Mike Fried

Abstract

Protein-induced DNA bending is of importance in the formation of complex nucleoprotein assemblies such as those involved in the initiation of DNA replication or transcription initiation. We have compared the DNA bending characteristics of the Escherichla coli cyclic AMP receptor protein (CRP or CAP), an archetypal DNA bending protein, to those of TFIID, the eukaryotic TATA-element binding transcription factor. By altering the helical phasing between a CRP binding site and the E.colimelR promoter we have mapped a DNA sequence-directed bend in the downstream region of the promoter. This intrinsic DNA bend may be important in the regulation of the melR promoter by CRP in vivo . Gel retardation assays and DNAse I footprinting show that human TFIID binds to the melR promoter -10 region. Taking advantage of this fact, and using the CRP-induced DNA bend as a standard, we have employed phase sensitive detection to show that the DNA bend angle induced by TFIID is far less than that induced by CRP. Further evidence to support this conclusion comes from a comparison of the relative mobilities of CRP-DNA and TFIID-DNA complexes. These results place limits on the role of any DNA bending induced by TFIID alone in the initiation of transcription.

Journal Article Type Article
Publication Date Jul 11, 1992
Journal Nucleic Acids Research
Print ISSN 0305-1048
Publisher Oxford University Press
Peer Reviewed Peer Reviewed
Volume 20
Issue 13
Pages 3391-3396
Institution Citation Gaston, K., Bell, A., Busby, S., & Fried, M. (1992). A comparison of the DNA bending properties of the DNA binding proteins CRP and TFIID. Nucleic Acids Research, 20(13), 3391-3396. doi:10.1093/nar/20.13.3391
DOI https://doi.org/10.1093/nar/20.13.3391
Publisher URL https://academic.oup.com/nar/article-abstract/20/13/3391/2383426?redirectedFrom=fulltext

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