The histone 3 lysine 36 methyltransferase, SET2, is involved in transcriptional elongation.

Existing evidence indicates that SET2, the histone 3 lysine 36 methyltransferase of Saccharomyces cerevisiae, is a transcriptional repressor. Here we show by five main lines of evidence that SET2 is involved in transcriptional elongation. First, most, if not all, subunits of the RNAP II holoenzyme co-purify with SET2. Second, all of the co-purifying RNAP II subunit, RPO21, was phosphorylated at serines 5 and 2 of the C-terminal domain (CTD) tail, indicating that the SET2 association is specific to either the elongating or SSN3 repressed forms (or both) of RNAP II. Third, the association of SET2 with CTD phosphorylated RPO21 remained in the absence of ssn3. Fourth, in the absence of ssn3, mRNA production from gal1 required SET2. Fifth, SET2 was detected on gal1 by in vivo crosslinking after, but not before, the induction of transcription. Similarly, SET2 physically associated with the transcribed region of pdr5 but was not detected on gal1 or pdr5 promoter regions. Since SET2 is also a histone methyltransferase, these results suggest a role for histone 3 lysine 36 methylation in transcriptional elongation.

[1]  Stuart L. Schreiber,et al.  Active genes are tri-methylated at K4 of histone H3 , 2002, Nature.

[2]  J. Greenblatt,et al.  Regulation of transcription elongation by phosphorylation. , 2002, Biochimica et biophysica acta.

[3]  Brian D. Strahl,et al.  Role of Histone H3 Lysine 9 Methylation in Epigenetic Control of Heterochromatin Assembly , 2001, Science.

[4]  R. Berezney,et al.  Growth-related Changes in Phosphorylation of Yeast RNA Polymerase II* , 1998, The Journal of Biological Chemistry.

[5]  Steven P. Gygi,et al.  Association of the Histone Methyltransferase Set2 with RNA Polymerase II Plays a Role in Transcription Elongation* , 2002, The Journal of Biological Chemistry.

[6]  M. Carlson,et al.  Cyclin-dependent protein kinase and cyclin homologs SSN3 and SSN8 contribute to transcriptional control in yeast. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Andrew J. Bannister,et al.  Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain , 2001, Nature.

[8]  R. Kornberg,et al.  A trithorax-group complex purified from Saccharomyces cerevisiae is required for methylation of histone H3 , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[9]  R. Young,et al.  Temporal regulation of RNA polymerase II by Srb10 and Kin28 cyclin-dependent kinases. , 1998, Molecular cell.

[10]  N. Thompson,et al.  Purification of eukaryotic RNA polymerase II by immunoaffinity chromatography. Elution of active enzyme with protein stabilizing agents from a polyol-responsive monoclonal antibody. , 1990, The Journal of biological chemistry.

[11]  T. Jenuwein,et al.  The many faces of histone lysine methylation. , 2002, Current opinion in cell biology.

[12]  C. Peterson,et al.  Purification and biochemical properties of yeast SWI/SNF complex. , 1999, Methods in enzymology.

[13]  Nevan J. Krogan,et al.  COMPASS, a Histone H3 (Lysine 4) Methyltransferase Required for Telomeric Silencing of Gene Expression* , 2002, The Journal of Biological Chemistry.

[14]  R. Paro,et al.  Gene regulation: Cycling silence , 2001, Nature.

[15]  Karl Mechtler,et al.  Methylation of histone H3 lysine 9 creates a binding site for HP1 proteins , 2001, Nature.

[16]  A Correspondent in Cell Biology,et al.  Gene Regulation , 1967, Nature.

[17]  P. Grant,et al.  Set2 Is a Nucleosomal Histone H3-Selective Methyltransferase That Mediates Transcriptional Repression , 2002, Molecular and Cellular Biology.

[18]  M Wilm,et al.  The S. cerevisiae SET3 complex includes two histone deacetylases, Hos2 and Hst1, and is a meiotic-specific repressor of the sporulation gene program. , 2001, Genes & development.

[19]  Assen Roguev,et al.  Deciphering Protein Complexes and Protein Interaction Networks by Tandem Affinity Purification and Mass Spectrometry , 2002, Molecular & Cellular Proteomics.

[20]  Michael R. Green,et al.  Dissecting the Regulatory Circuitry of a Eukaryotic Genome , 1998, Cell.

[21]  Ken-ichi Noma,et al.  Transitions in Distinct Histone H3 Methylation Patterns at the Heterochromatin Domain Boundaries , 2001, Science.

[22]  R. Kornberg,et al.  Quantitation of the RNA Polymerase II Transcription Machinery in Yeast* , 2001, The Journal of Biological Chemistry.

[23]  J. Greenblatt,et al.  Opposing effects of Ctk1 kinase and Fcp1 phosphatase at Ser 2 of the RNA polymerase II C-terminal domain. , 2001, Genes & development.

[24]  Stuart L. Schreiber,et al.  Methylation of histone H3 Lys 4 in coding regions of active genes , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[25]  T. Richmond,et al.  Crystal structure of the nucleosome core particle at 2.8 Å resolution , 1997, Nature.

[26]  B. Séraphin,et al.  A generic protein purification method for protein complex characterization and proteome exploration , 1999, Nature Biotechnology.

[27]  Rein Aasland,et al.  The Saccharomyces cerevisiae Set1 complex includes an Ash2 homologue and methylates histone 3 lysine 4 , 2001, The EMBO journal.

[28]  J. Archambault,et al.  Genetic interaction between transcription elongation factor TFIIS and RNA polymerase II , 1992, Molecular and cellular biology.

[29]  M. Grunstein,et al.  Mapping DNA interaction sites of chromosomal proteins using immunoprecipitation and polymerase chain reaction. , 1999, Methods in enzymology.

[30]  R. Young,et al.  Negative regulation of Gcn4 and Msn2 transcription factors by Srb10 cyclin-dependent kinase. , 2001, Genes & development.

[31]  Danny Reinberg,et al.  RNA polymerase II elongation through chromatin , 2000, Nature.

[32]  B. Séraphin,et al.  New constructs and strategies for efficient PCR‐based gene manipulations in yeast , 1998, Yeast.