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Molecular and Cellular Biology, March 2008, p. 1883-1891, Vol. 28, No. 6
0270-7306/08/$08.00+0     doi:10.1128/MCB.01714-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

The Rpb4 Subunit of RNA Polymerase II Contributes to Cotranscriptional Recruitment of 3' Processing Factors{triangledown} ,{dagger}

Vanessa M. Runner, Vladimir Podolny, and Stephen Buratowski*

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, Massachusetts 02115

Received 18 September 2007/ Returned for modification 18 October 2007/ Accepted 28 December 2007

The RNA polymerase II enzyme from the yeast Saccharomyces cerevisiae is a complex of 12 subunits, Rpb1 to Rpb12. Crystal structures of the full complex show that the polymerase consists of two separable components, a 10-subunit core including the catalytic active site and a heterodimer of the Rpb4 and Rpb7 subunits. To characterize the role of the Rpb4/7 heterodimer during transcription in vivo, chromatin immunoprecipitation was used to examine an rpb4{Delta} strain for effects on the behavior of the core polymerase as well as recruitment of other protein factors involved in transcription. Rpb4/7 cross-links throughout transcribed regions. Loss of Rpb4 results in a reduction of RNA polymerase II levels near 3' ends of multiple mRNA genes as well as a decreased association of 3'-end processing factors. Furthermore, loss of Rpb4 results in altered polyadenylation site usage at the RNA14 gene. Together, these results indicate that Rpb4 contributes to proper cotranscriptional 3'-end processing in vivo.

* Corresponding author. Mailing address: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115. Phone: (617) 432-0696. Fax: (617) 738-0516. E-mail: SteveB{at}hms.harvard.edu

{triangledown} Published ahead of print on 14 January 2008.

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

Molecular and Cellular Biology, March 2008, p. 1883-1891, Vol. 28, No. 6
0270-7306/08/$08.00+0     doi:10.1128/MCB.01714-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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