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Molecular and Cellular Biology, April 2009, p. 2308-2321, Vol. 29, No. 8
0270-7306/09/$08.00+0     doi:10.1128/MCB.01841-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Sub1 Functions in Osmoregulation and in Transcription by both RNA Polymerases II and III{triangledown} ,{dagger}

Emanuel Rosonina,1 Ian M. Willis,2 and James L. Manley1*

Department of Biological Sciences, Columbia University, New York, New York 10027,1 Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York 104612

Received 3 December 2008/ Returned for modification 12 January 2009/ Accepted 29 January 2009

Sub1 is implicated in transcriptional activation, elongation, and mRNA 3'-end formation in budding yeast. To gain more insight into its function, we performed a synthetic genetic array screen with SUB1 that uncovered genetic interactions with genes involved in the high-osmolarity glycerol (HOG) osmoresponse pathway. We find that Sub1 and the HOG pathway are redundant for survival in moderate osmolarity. Chromatin immunoprecipitation analysis shows that Sub1 is recruited to osmoresponse gene promoters during osmotic shock and is required for full recruitment of TBP, TFIIB, and RNA polymerase II (RNAP II) at a subset of these genes. Furthermore, we detect Sub1 at the promoter of every constitutively transcribed RNAP II and, unexpectedly, at every RNAP III gene tested, but not at the RNAP I-transcribed ribosomal DNA promoter. Significantly, deletion of SUB1 reduced levels of promoter-associated RNAP II or III at these genes, but not TBP levels. Together these data suggest that, in addition to a general role in polymerase recruitment at constitutive RNAP II and RNAP III genes, during osmotic shock, Sub1 facilitates osmoresponse gene transcription by enhancing preinitiation complex formation.

* Corresponding author. Mailing address: Department of Biological Sciences, Columbia University, New York, NY 10027. Phone: (212) 854-4647. Fax: (212) 865-8246. E-mail: jlm2{at}columbia.edu

{triangledown} Published ahead of print on 9 February 2009.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, April 2009, p. 2308-2321, Vol. 29, No. 8
0270-7306/09/$08.00+0     doi:10.1128/MCB.01841-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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