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J. Biol. Chem., Vol. 276, Issue 46, 43056-43064, November 16, 2001
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From the Chemical Pathology Unit, Sir William Dunn School of
Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom
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 proximal sequence elements
(PSEs) 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. 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 TATA-binding protein (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 adenovirus major late promoter (AdMLP). 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.
BRFU, a TFIIB-like Factor, Is Directly Recruited to the
TATA-box of Polymerase III Small Nuclear RNA Gene Promoters through
Its Interaction with TATA-binding Protein*
abart
§ and¶
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported by MRC Senior Fellowship No. G117/309.
§
To whom correspondence may be addressed. Tel.: 44-1-865-275-608;
Fax: 44-1-865-275-556; E-mail: pcabart@molbiol.ox.ac.uk.
¶
To whom correspondence may be addressed. Tel.:
44-1-865-275-608; Fax: 44-1-865-275-556; E-mail:
smurphy@molbiol.ox.ac.uk.
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