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Nucleic Acids Research Advance Access originally published online on October 6, 2006
Nucleic Acids Research 2006 34(19):5585-5593; doi:10.1093/nar/gkl534
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Nucleic Acids Research, 2006, Vol. 34, No. 19 5585-5593
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

The Saccharomyces cerevisiae RNA polymerase III recruitment factor subunits Brf1 and Bdp1 impose a strict sequence preference for the downstream half of the TATA box

Nick D. Tsihlis and Anne Grove*

Department of Biological Sciences, Louisiana State University Baton Rouge, LA 70803, USA

*To whom correspondence should be addressed. Tel: +1 225 578 5148; Fax: +1 225 578 8790; Email: agrove{at}lsu.edu

Received June 5, 2006. Revised July 7, 2006. Accepted July 10, 2006.

Association of the TATA-binding protein (TBP) with its cognate site within eukaryotic promoters is key to accurate and efficient transcriptional initiation. To achieve recruitment of Saccharomyces cerevisiae RNA polymerase III, TBP is associated with two additional factors, Brf1 and Bdp1, to form the initiation factor TFIIIB. Previous data have suggested that the structure or dynamics of the TBP–DNA complex may be altered upon entry of Brf1 and Bdp1 into the complex. We show here, using the altered specificity TBP mutant TBPm3 and an iterative in vitro selection assay, that entry of Brf1 and Bdp1 into the complex imposes a strict sequence preference for the downstream half of the TATA box. Notably, the selected sequence (TGTAAATA) is a perfect match to the TATA box of the RNA polymerase III-transcribed U6 small nuclear RNA (SNR6) gene. We suggest that the selected T•A base pair step at the downstream end of the 8 bp TBP site may provide a DNA flexure that promotes TFIIIB-DNA complex formation.

Present address: N. D. Tsihlis, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA


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