An immunoaffinity purified Schizosaccharomyces pombe TBP-containing complex directs correct initiation of the S-pombe rRNA gene promoter

被引：6

作者：

Chen, L ^{[1
]}

Guo, A ^{[1
]}

Pape, L ^{[1
]}

机构：

[1] NYU, DEPT CHEM, NEW YORK, NY 10003 USA

来源：

NUCLEIC ACIDS RESEARCH | 1997年 / 25卷 / 08期

基金：

美国国家科学基金会;

关键词：

D O I：

10.1093/nar/25.8.1633

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The multi-protein complex SL1, containing TBP, which is essential for RNA polymerase I catalyzed transcription, has been analyzed in fission yeast. It was immunopurified based on association of component subunits with epitope-tagged TBP. To enable this analysis, a strain of Schizosaccharomyces pombe was created where the only functional TBP coding sequences were those of FLAG-TBP. RNA polymerase I transcription components were fractionated from this strain and the TBP-associated polypeptides were subsequently immunopurified together with the epitope-tagged TBP. An assessment of the activity of this candidate SL1 complex was undertaken cross-species. This fission yeast TBP-containing complex displays two activities in redirecting transcriptional initiation of an S.pombe rDNA gene promoter cross-species in Saccharomyces cerevisiae transcription reactions: it both blocks an incorrect transcriptional start site at +7 and directs initiation at the correct site for S.pombe rRNA synthesis. This complex is essential for accurate initiation of the S.pombe rRNA gene: rRNA synthesis is reconstituted when this S.pombe TBP-containing complex is combined with a S.pombe fraction immunodepleted of TBP.

引用

页码：1633 / 1640

页数：8

共 55 条

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Ausubel FM., 1994, Curr. Protoc. Mol. Biol

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