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Molecular and Cellular Biology, October 2002, p. 6697-6705, Vol. 22, No. 19
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.19.6697-6705.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Core Promoter-Dependent TFIIB Conformation and a Role for TFIIB Conformation in Transcription Start Site Selection

Jennifer A. Fairley,2,{dagger} Rachel Evans,1,2 Nicola A. Hawkes,2,{ddagger} and Stefan G. E. Roberts1,2*

School of Biological Sciences, University of Manchester, Manchester M13 9PT,1 Wellcome Trust Biocentre, University of Dundee, Dundee DD1 5EH2

Received 21 May 2002/ Accepted 21 June 2002

The general transcription factor TFIIB plays a central role in the selection of the transcription initiation site. The mechanisms involved are not clear, however. In this study, we analyze core promoter features that are responsible for the susceptibility to mutations in TFIIB and cause a shift in the transcription start site. We show that TFIIB can modulate both the 5' and 3' parameters of transcription start site selection in a manner dependent upon the sequence of the initiator. Mutations in TFIIB that cause aberrant transcription start site selection concentrate in a region that plays a pivotal role in modulating TFIIB conformation. Using epitope-specific antibody probes, we show that a TFIIB mutant that causes aberrant transcription start site selection assembles at the promoter in a conformation different from that for wild-type TFIIB. In addition, we uncover a core promoter-dependent effect on TFIIB conformation and provide evidence for novel sequence-specific TFIIB promoter contacts.

* Corresponding author. Mailing address: School of Biological Sciences, G.186 Stopford Building, University of Manchester, Oxford Road, Manchester M13 9PT, United Kingdom. Phone: 0161 275 5758. Fax: 0161 275 5600. E-mail: stefan.roberts{at}man.ac.uk.

{dagger} Present address: Division of Biochemistry and Molecular Biology, Davidson Building, University of Glasgow, Glasgow G12 8QQ, United Kingdom.

{ddagger} Present address: Clare Hall Laboratories, ICRF, South Mimms, Hertfordshire EN6 3LD, United Kingdom.

Molecular and Cellular Biology, October 2002, p. 6697-6705, Vol. 22, No. 19
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.19.6697-6705.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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