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The transcriptional coactivator sub1 has a role in transcription elongation

AuthorsGarcía, Alicia ; Rosonina, Emanuel; Manley, James L.; Calvo, Olga
Issue Date2008
Citation8th EMBL Transcription Meeting (2008)
AbstractWe have shown that the transcriptional coactivator SubI is recruited to promoters and remains chromatin-associaled along the length of several yeast genes during transcription elongation. We also found allele-specific interactions between SUBI and the RNAP II kinase. Kin28 and phosphatase, Fcpl. Furthemore, cells lacking SubI display decreased accumulalion of RNAP II phosphatase, Fcp I, altered RNAP II phosphorylation and decreased crosslinking of RNAP II lo transcribed genes. These results strongly suggested thal SubI has a role in Transcriplion elongation. The carboxy-tenninal domain of thc RNAP ll largest subunit (CTD) is phosphorylated by the action or several eyelin-dependent kinases. In Saccharomyces cerevisiae, there are at leasl four CTD kinases with different roles in the transcription cycle: Srb I0 (preinitiation), Kin28 (initiation), Ctk I and Burl (elongalion). To further investigate the role of SubI in CTD phosphorylation and in the transcription cycle, we performed genetic studies with different CTD kinase mutant strains: srbI0', burl-23, burI-l and ctkI. SrbI0 phosphorylation inhibits RNAP II entry into the pre-initiation complex (PlC). We find that the growth defect displayed by srbI0' cells is partially suppressed by SUBI deletion, and dramatically enhanced by SUBI overexpression, implicating Subl in promoting PIC formation. For burl mutants, SUBI deletion in bur1/-23 and burI-l cells enhanced their slow growth and caused sensitivity to high temperature and to 6-azauracil, an indicator of defects in elongation. Additionally, the growth defect seen in ctkl' cells is greatly enhanced in the absence of SUBl. We have also found that the subl' strain shows a decrease in elongation efficiency as determined by the GLAM assay, which measures gene length-dependent accumulation of mRNAs in vivo. Furthemore, in vitro kinase assays performed in wild type and subl' cells, indicate that Subl affects CTD-Ser5 phosphorylation by Kin28. Taken together, our data point to a role for Subl in the regulalion of transcription by RNAP ll at initialion and elongation, through regulation of CTD phosphorylation at multiple levels.
DescriptionResumen del póster presentado al 8th European Molecular Biology Laboratory (EMBL) Transcription Meeting, celebrado en Heidelberg (Alemania) del 23 al 27 de agosto de 2008.
Appears in Collections:(IMB) Comunicaciones congresos
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