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Molecular and Cellular Biology, September 2001, p. 5790-5796, Vol. 21, No. 17
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.17.5790-5796.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Interaction of the Srb10 Kinase with Sip4, a Transcriptional Activator of Gluconeogenic Genes in Saccharomyces cerevisiae

Olivier Vincent,1 Sergei Kuchin,1 Seung-Pyo Hong,1 Robert Townley,2 Valmik K. Vyas,2 and Marian Carlson1,2,*

Departments of Genetics and Development and Microbiology1 and Integrated Program in Cellular Biology, Molecular Biology and Biophysical Studies,2 Columbia University, New York, New York 10032

Received 7 March 2001/Returned for modification 23 April 2001/Accepted 13 June 2001

Sip4 is a Zn2Cys6 transcriptional activator that binds to the carbon source-responsive elements of gluconeogenic genes in Saccharomyces cerevisiae. The Snf1 protein kinase interacts with Sip4 and regulates its phosphorylation and activator function in response to glucose limitation; however, evidence suggested that another kinase also regulates Sip4. Here we examine the role of the Srb10 kinase, a component of the RNA polymerase II holoenzyme that has been primarily implicated in transcriptional repression but also positively regulates Gal4. We show that Srb10 is required for phosphorylation of Sip4 during growth in nonfermentable carbon sources and that the catalytic activity of Srb10 stimulates the ability of LexA-Sip4 to activate transcription of a reporter. Srb10 and Sip4 coimmunoprecipitate from cell extracts and interact in two-hybrid assays, suggesting that Srb10 regulates Sip4 directly. We also present evidence that the Srb10 and Snf1 kinases interact with different regions of Sip4. These findings support the view that the Srb10 kinase not only plays negative roles in transcriptional control but also has broad positive roles during growth in carbon sources other than glucose.

* Corresponding author. Mailing address: Columbia University, 701 W. 168th St., HSC 922, New York, NY 10032. Phone: (212) 305-6314. Fax: (212) 305-1741. E-mail: mbc1{at}columbia.edu.

Molecular and Cellular Biology, September 2001, p. 5790-5796, Vol. 21, No. 17
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.17.5790-5796.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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