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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.

Original publication

DOI

10.1074/jbc.M108515200

Type

Journal article

Journal

J Biol Chem

Publication Date

16/11/2001

Volume

276

Pages

43056 - 43064

Keywords

Amino Acids, Base Sequence, Cell Nucleus, DNA Polymerase III, DNA-Binding Proteins, Deoxyribonuclease I, Glutathione Transferase, Histidine, Humans, Models, Biological, Molecular Sequence Data, Mutation, Promoter Regions, Genetic, Protein Binding, Protein Structure, Tertiary, RNA, Recombinant Fusion Proteins, TATA Box, TATA-Box Binding Protein, Transcription Factor TFIIIB, Transcription Factors, Transcription, Genetic