Open AccessAn Immunoaffinity Purified Schizosaccharomyces Pombe TBP-containing Complex Directs Correct Initiation of the S.pombe rRNA Gene Promoter
Author(s) -
Ling Chen,
Ailan Guo,
Louise Pape
Publication year - 1997
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/25.8.1633
Subject(s) - schizosaccharomyces pombe , biology , schizosaccharomyces , transcription preinitiation complex , transcription (linguistics) , ribosomal rna , rna polymerase ii , yeast , saccharomyces cerevisiae , gene , microbiology and biotechnology , rna polymerase i , genetics , rna polymerase , rna , promoter , gene expression , linguistics , philosophy
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.