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Molecular and Cellular Biology, April 2002, p. 2788-2798, Vol. 22, No. 8
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.8.2788-2798.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Evidence that TAF-TATA Box-Binding Protein Interactions Are Required for Activated Transcription in Mammalian Cells

Lisa S. Martel,1 Helen J. Brown,2 and Arnold J. Berk1,3*

Department of Microbiology, Immunology, and Molecular Genetics,1 Department of Molecular Pharmacology,2 Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California3

Received 27 August 2001/ Returned for modification 8 October 2001/ Accepted 20 December 2001

Surfaces of human TATA box-binding protein (hsTBP) required for activated transcription in vivo were defined by constructing a library of surface residue substitution mutations and assaying them for their ability to support activated transcription in transient-transfection assays. In earlier work, three regions were identified where mutations inhibited activated transcription without interfering with TATA box DNA binding. One region is on the upstream surface of the N-terminal TBP repeat with respect to the direction of transcription and corresponds to the TBP surface that interacts with TFIIA. A second region on the stirrup of the C-terminal TBP repeat corresponds to the TFIIB-binding surface. Here we report that the third region where mutations inhibit activated transcription in mammalian cells, the convex surface of the N-terminal repeat, corresponds to a surface on TBP that interacts with hsTAF1, the major scaffold subunit of TFIID. Since mutations at the center of the hsTAF1-interacting region inhibit the ability of the protein to support activated transcription in vivo, these results are consistent with the conclusion that an interaction between hsTBP and TAFIIs is required for activated transcription in mammalian cells.

* Corresponding author. Mailing address: Paul D. Boyer Hall, 611 Charles Young Dr. E, Box 951570, Los Angeles, CA 90095-1570. Phone: (310) 206-6298. Fax: (310) 206-7286. E-mail: berk{at}mbi.ucla.edu.

Molecular and Cellular Biology, April 2002, p. 2788-2798, Vol. 22, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.8.2788-2798.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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