Yeast Genetics and Molecular Biology 2000
University of Washington
Seattle, Washington USA
July 2000


Name: Costa, Patrick J.
Mailing Address: Dept. of Biological Sciences, University of Pittsburgh, 269 Crawford Hall, Pittsburgh, PA 15260, USA
Email Address: pjcst25+@pitt.edu
Phone & FAX numbers: (412) 624-6992 & (412) 624-4759

#045

Evidence that the Saccharomyces cerevisiae Rtf1 protein regulates transcription elongation.
Patrick J. Costa, Karen M. Arndt
Dept. of Biological Sciences, University of Pittsburgh, 269 Crawford Hall, Pittsburgh, PA 15260, USA

RTF1 was originally identified in a search for suppressors of a TBP altered specificity mutant. To elucidate the function of Rtf1, a synthetic lethal screen with an rtf1 null allele was performed. The screen identified mutations in nine genes. The cloning of six of the genes showed that they define two classes. SWI4 and SWI6 encode activators of various cell-cycle regulated promoters, while SRB5, CTK1, FCP1, and POB3 have more global roles in transcription. Specifically, SRB5, CTK1, and FCP1 directly affect the phosphorylation state of the C-terminal domain (CTD) of RNA polymerase II. The isolation of mutations in these genes argues that Rtf1 function is essential when CTD phosphorylation is abnormal. Because the extent of CTD phosphorylation correlates strongly with progression from transcription initiation to elongation, our results also suggest that Rtf1 may be important for this transition, particularly in the context of chromatin. Interestingly, we uncovered a POB3 mutation in our screen. Pob3 shares similarity with HMG1-like proteins and forms a complex with Cdc68/Spt16. The human counterparts of Pob3 and Cdc68/Spt16 form a nucleosome-specific transcription elongation factor. A role for Rtf1 in elongation is further supported by interactions between RTF1 and genes encoding several known elongation factors. In addition, rtfl null mutations confer sensitivity to 6-azauracil and mycophenolic acid, phenotypes associated with defects in transcription elongation.


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