An Immunoaffinity Purified Schizosaccharomyces Pombe TBP-containing Complex Directs Correct Initiation of the S.pombe RRNA Gene Promoter

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1997 Apr 15

PMID 9092673

Citations 2

Authors

L Chen

A Guo

L Pape

Affiliations

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Abstract

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.

Citing Articles

Characterization of the fission yeast ribosomal DNA binding factor: components share homology with Upstream Activating Factor and with SWI/SNF subunits.

Liu M, Guo A, Boukhgalter B, Van Den Heuvel K, Tripp M, Pape L Nucleic Acids Res. 2002; 30(24):5347-59.

PMID: 12490702 PMC: 140063. DOI: 10.1093/nar/gkf683.

Widespread use of TATA elements in the core promoters for RNA polymerases III, II, and I in fission yeast.

Hamada M, Huang Y, Lowe T, Maraia R Mol Cell Biol. 2001; 21(20):6870-81.

PMID: 11564871 PMC: 99864. DOI: 10.1128/MCB.21.20.6870-6881.2001.

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