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A more recent version of this article appeared on November 9, 2001
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M108515200v1
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Papers In Press, published online ahead of print September 19, 2001
J. Biol. Chem, 10.1074/jbc.M108515200
Submitted on September 5, 2001
Revised on September 19, 2001
Accepted on September 18, 2001

BRFU, a TFIIB-like factor, is directly recruited to the TATA-box of Pol III snRNA gene promoters through its interaction with TBP

Pavel Cabart and Shona Murphy

Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE

Corresponding Author: pcabart{at}molbiol.ox.ac.uk

The human snRNA genes transcribed by RNA polymerase II (Pol II) and III (Pol III) have different core promoter elements. Both gene types contain similar PSE elements, but differ in the absence (Pol II) or presence (Pol III) of a TATA-box, which, together with the PSE, determines the assembly of a Pol III-specific pre-initiation complex (PIC). BRFU is a factor exclusively required for transcription of the Pol III-type snRNA genes. We report that recruitment of BRFU to the TATA-box of these promoters is TBP-dependent. BRFU in turn stabilizes TBP on TATA-containing template and extends the TBP footprint both upstream and downstream of the TATA element. The core domain of TBP is sufficient for BRFU:TBP:DNA complex formation and for interaction with BRFU off the template. We have mapped amino-acid residues within TBP and domains of BRFU that mediate this interaction. BRFU has no specificity for sequences flanking the TATA-box and also forms a stable complex on the TATA-box of the Pol II-specific AdML promoter. Furthermore, Pol III-type transcription can initiate from an snRNA gene promoter containing an AdMLP TATA-box and flanking sequences. Therefore, the polymerase recruitment is not simply determined by the sequence of the TATA-box and immediate flanking sequences.


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