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Molecular and Cellular Biology, June 2003, p. 4207-4218, Vol. 23, No. 12
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.12.4207-4218.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Methylation of Histone H3 by Set2 in Saccharomyces cerevisiae Is Linked to Transcriptional Elongation by RNA Polymerase II

Nevan J. Krogan,1,2,3 Minkyu Kim,4 Amy Tong,1,2 Ashkan Golshani,1 Gerard Cagney,1,2,3 Veronica Canadien,5 Dawn P. Richards,5 Bryan K. Beattie,5 Andrew Emili,1,2,3 Charles Boone,1,2 Ali Shilatifard,6 Stephen Buratowski,4 and Jack Greenblatt1,2,3*

Banting and Best Department of Medical Research,1 Toronto Yeast Proteomics Organization, University of Toronto, Toronto, Ontario, Canada M5G 1L6,3 Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario, Canada M5S 1A8,2 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115,4 Affinium Pharmaceuticals, Toronto, Ontario, Canada, M5J 1V6,5 Department of Biochemistry and Cancer Center, St. Louis University School of Medicine, St. Louis, Missouri 631046

Received 13 January 2003/ Returned for modification 27 February 2003/ Accepted 20 March 2003

Set2 methylates Lys36 of histone H3. We show here that yeast Set2 copurifies with RNA polymerase II (RNAPII). Chromatin immunoprecipitation analyses demonstrated that Set2 and histone H3 Lys36 methylation are associated with the coding regions of several genes that were tested and correlate with active transcription. Both depend, as well, on the Paf1 elongation factor complex. The C terminus of Set2, which contains a WW domain, is also required for effective Lys36 methylation. Deletion of CTK1, encoding an RNAPII CTD kinase, prevents Lys36 methylation and Set2 recruitment, suggesting that methylation may be triggered by contact of the WW domain or C terminus of Set2 with Ser2-phosphorylated CTD. A set2 deletion results in slight sensitivity to 6-azauracil and much less ß-galactosidase produced by a reporter plasmid, resulting from a defect in transcription. In synthetic genetic array (SGA) analysis, synthetic growth defects were obtained when a set2 deletion was combined with deletions of all five components of the Paf1 complex, the chromodomain elongation factor Chd1, the putative elongation factor Soh1, the Bre1 or Lge1 components of the histone H2B ubiquitination complex, or the histone H2A variant Htz1. SET2 also interacts genetically with components of the Set1 and Set3 complexes, suggesting that Set1, Set2, and Set3 similarly affect transcription by RNAPII.

* Corresponding author. Mailing address: Banting and Best Department of Medical Research, University of Toronto, 112 College St., Toronto ON, Canada M5G 1L6. Phone: (416) 978-4141. Fax: (416) 978-8528. E-mail: jack.greenblatt{at}utoronto.ca.

Molecular and Cellular Biology, June 2003, p. 4207-4218, Vol. 23, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.12.4207-4218.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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