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Eukaryotic Cell, August 2005, p. 1446-1454, Vol. 4, No. 8
1535-9778/05/$08.00+0     doi:10.1128/EC.4.8.1446-1454.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Histone H3 K36 Methylation Is Associated with Transcription Elongation in Schizosaccharomyces pombe

Stephanie A. Morris,1 Yoichiro Shibata,1 Ken-ichi Noma,2 Yuko Tsukamoto,2 Erin Warren,1 Brenda Temple,1 Shiv I. S. Grewal,2 and Brian D. Strahl1*

Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599,1 Laboratory of Molecular Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 208922

Received 23 March 2005/ Accepted 29 May 2005

Set2 methylation of histone H3 at lysine 36 (K36) has recently been shown to be associated with RNA polymerase II (Pol II) elongation in Saccharomyces cerevisiae. However, whether this modification is conserved and associated with transcription elongation in other organisms is not known. Here we report the identification and characterization of the Set2 ortholog responsible for K36 methylation in the fission yeast Schizosaccharomyces pombe. We find that similar to the budding yeast enzyme, S. pombe Set2 is also a robust nucleosome-selective H3 methyltransferase that is specific for K36. Deletion of the S. pombe set2+ gene results in complete abolishment of K36 methylation as well as a slow-growth phenotype on plates containing synthetic medium. These results indicate that Set2 is the sole enzyme responsible for this modification in fission yeast and is important for cell growth under stressed conditions. Using the chromatin immunoprecipitation assay, we demonstrate that K36 methylation in S. pombe is associated with the transcribed regions of Pol II-regulated genes and is devoid in regions that are not transcribed by Pol II. Consistent with a role for Set2 in transcription elongation, we find that S. pombe Set2 associates with the hyperphosphorylated form of Pol II and can fully rescue K36 methylation and Pol II interaction in budding yeast cells deleted for Set2. These results, along with our finding that K36 methylation is highly conserved among eukaryotes, imply a conserved role for this modification in the transcription elongation process.

* Corresponding author. Mailing address: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599. Phone: 919-843-3896. Fax: 919-966-2852. E-mail: brian_strahl{at}med.unc.edu.

Eukaryotic Cell, August 2005, p. 1446-1454, Vol. 4, No. 8
1535-9778/05/$08.00+0     doi:10.1128/EC.4.8.1446-1454.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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