Transcription of the Schizosaccharomyces pombe gene cdc18+: roles of MCB elements and the DSC1 complex.
In Schizosaccharomyces pombe, commitment to a round of DNA synthesis and entry into the cell cycle are dependent on the function of genes that are transcribed periodically during the cell cycle. Activation of these genes prior to S phase is primarily controlled through cis-acting elements known as MluI Cell-cycle Boxes, or MCBs, and by a family of transcription factors, including Cdc10, Res1, Res2 and Rep2. These transcription factors are also known to be present in a complex, DSC1, that binds to the promoters of pre-S genes. We have demonstrated that within the promoter of cdc18+, a representative pre-S gene, the orientation and spacing of MCBs are crucial for activation and cell-cycle dependence. To our surprise, electrophoretic mobility shift assays showed a highly active mutant form of the promoter, which alters the spacing of the MCB elements, does not bind DSC1 but does bind a higher mobility complex. The binding of this second complex is not dependent on Cdc10 or the Res/Rep proteins. We conclude that, DSC1 binding does not correlate with cell-cycle dependent transcriptional activation, and the higher mobility species may represent a novel transcriptional activation complex that is also likely to function in pre-S transcription.
Jackson
William T
WT
Department of Microbiology and Immunology, Stanford University Medical Center, Fairchild D315, 299 Campus Drive, Stanford, CA 94305, United States. wjackson@stanford.edu
Martin
G Steven
GS
D018797:Cell Cycle Proteins / Q000235:genetics*; D024202:Electrophoretic Mobility Shift Assay; D005800:Genes, Fungal; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
D018797:Cell Cycle Proteins; C489911:DSC1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C082462:cdc18 protein, S pombe