title: Structure/Function Analysis of Recurrent Mutations in SETD2 Protein Reveals a Critical and Conserved Role for a SET Domain Residue in Maintaining Protein Stability and Histone H3 Lys-36 Trimethylation.
abstract: The yeast Set2 histone methyltransferase is a critical enzyme that plays a number of key roles in gene transcription and DNA repair. Recently, the human homologue, SETD2, was found to be recurrently mutated in a significant percentage of renal cell carcinomas, raising the possibility that the activity of SETD2 is tumor-suppressive. Using budding yeast and human cell line model systems, we examined the functional significance of two evolutionarily conserved residues in SETD2 that are recurrently mutated in human cancers. Whereas one of these mutations (R2510H), located in the Set2 Rpb1 interaction domain, did not result in an observable defect in SETD2 enzymatic function, a second mutation in the catalytic domain of this enzyme (R1625C) resulted in a complete loss of histone H3 Lys-36 trimethylation (H3K36me3). This mutant showed unchanged thermal stability as compared with the wild type protein but diminished binding to the histone H3 tail. Surprisingly, mutation of the conserved residue in Set2 (R195C) similarly resulted in a complete loss of H3K36me3 but did not affect dimethylated histone H3 Lys-36 (H3K36me2) or functions associated with H3K36me2 in yeast. Collectively, these data imply a critical role for Arg-1625 in maintaining the protein interaction with H3 and specific H3K36me3 function of this enzyme, which is conserved from yeast to humans. They also may provide a refined biochemical explanation for how H3K36me3 loss leads to genomic instability and cancer.
authors.affiliation: From the Department of Genetics, Curriculum in Genetics and Molecular Biology, the Lineberger Comprehensive Cancer Center, and.
authors.forename: Kathryn E
authors.initials: KE
authors.lastname: Hacker
authors.affiliation: From the Department of Genetics, Curriculum in Genetics and Molecular Biology, the Lineberger Comprehensive Cancer Center, and.
authors.forename: Catherine C
authors.initials: CC
authors.lastname: Fahey
authors.affiliation: the Lineberger Comprehensive Cancer Center, and the Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina 27599.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Shinsky
authors.affiliation: the Lineberger Comprehensive Cancer Center, and.
authors.forename: Yun-Chen J
authors.initials: YJ
authors.lastname: Chiang
authors.affiliation: From the Department of Genetics, Curriculum in Genetics and Molecular Biology, the Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina 27599.
authors.forename: Julia V
authors.initials: JV
authors.lastname: DiFiore
authors.affiliation: the Lineberger Comprehensive Cancer Center, and the Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina 27599.
authors.forename: Deepak Kumar
authors.initials: DK
authors.lastname: Jha
authors.affiliation: the Department of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Andy H
authors.initials: AH
authors.lastname: Vo
authors.affiliation: the Department of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Jordan A
authors.initials: JA
authors.lastname: Shavit
authors.affiliation: From the Department of Genetics, Curriculum in Genetics and Molecular Biology, the Lineberger Comprehensive Cancer Center, and the Department of Pediatrics, University of North Carolina, Chapel Hill, North Carolina 27514, and.
authors.forename: Ian J
authors.initials: IJ
authors.lastname: Davis
authors.affiliation: From the Department of Genetics, Curriculum in Genetics and Molecular Biology, the Lineberger Comprehensive Cancer Center, and the Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina 27599, brian_strahl@med.unc.edu.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Strahl
authors.affiliation: From the Department of Genetics, Curriculum in Genetics and Molecular Biology, the Lineberger Comprehensive Cancer Center, and the Division of Hematology and Oncology, Department of Cancer Biology, Vanderbilt-Ingram Cancer Center, Nashville, Tennessee 37232 kimryn.rathmell@vanderbilt.edu.
authors.forename: W Kimryn
authors.initials: WK
authors.lastname: Rathmell
mesh_terms: D004795:Enzyme Stability / Q000235:genetics; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D006657:Histones; D009363:Neoplasm Proteins; D029701:Saccharomyces cerevisiae Proteins; D008780:Methyltransferases; C448996:Set2 protein, S cerevisiae; D011495:Histone-Lysine N-Methyltransferase; C521636:SETD2 protein, human
keywords: H3K36; SETD2; Set2; cancer; chromatin; enzyme; histone methylation; mutant