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Molecular and Cellular Biology, November 1999, p. 7610-7620, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A Human TATA Binding Protein-Related Protein with Altered DNA Binding Specificity Inhibits Transcription from Multiple Promoters and Activators

Paul A. Moore,1 Josef Ozer,2 Moreh Salunek,2 Gwenael Jan,3 Dennis Zerby,2 Susan Campbell,3 and Paul M. Lieberman2,*

Human Genome Sciences, Rockville, Maryland 208501; The Wistar Institute, Philadelphia, Pennsylvania 191042; and Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB9 1AS, Scotland3

Received 17 June 1999/Accepted 28 July 1999

The TATA binding protein (TBP) plays a central role in eukaryotic and archael transcription initiation. We describe the isolation of a novel 23-kDa human protein that displays 41% identity to TBP and is expressed in most human tissue. Recombinant TBP-related protein (TRP) displayed barely detectable binding to consensus TATA box sequences but bound with slightly higher affinities to nonconsensus TATA sequences. TRP did not substitute for TBP in transcription reactions in vitro. However, addition of TRP potently inhibited basal and activated transcription from multiple promoters in vitro and in vivo. General transcription factors TFIIA and TFIIB bound glutathione S-transferase-TRP in solution but failed to stimulate TRP binding to DNA. Preincubation of TRP with TFIIA inhibited TBP-TFIIA-DNA complex formation and addition of TFIIA overcame TRP-mediated transcription repression. TRP transcriptional repression activity was specifically reduced by mutations in TRP that disrupt the TFIIA binding surface but not by mutations that disrupt the TFIIB or DNA binding surface of TRP. These results suggest that TFIIA is a primary target of TRP transcription inhibition and that TRP may modulate transcription by a novel mechanism involving the partial mimicry of TBP functions.

* Corresponding author. Mailing address: The Wistar Institute, 3601 Spruce St., Philadelphia, PA 19104-4205. Phone: (215) 898-9491. Fax: (215) 898-0663. E-mail: lieberman{at}wista.wistar.upenn.edu.

Molecular and Cellular Biology, November 1999, p. 7610-7620, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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