title: Unraveling Site-Specific and Combinatorial Histone Modifications Using High-Resolution Mass Spectrometry in Histone Deacetylase Mutants of Fission Yeast.
abstract: Histone deacetylases (HDACs) catalyze the removal of acetylation marks from lysine residues on histone and nonhistone substrates. Their activity is generally associated with essential cellular processes such as transcriptional repression and heterochromatin formation. Interestingly, abnormal activity of HDACs has been reported in various types of cancers, which makes them a promising therapeutic target for cancer treatment. In the current study, we aim to understand the mechanisms underlying the function of HDACs using an in-depth quantitative analysis of changes in histone acetylation levels in Schizosaccharomyces pombe (S. pombe) lacking major HDAC activities. We employed a targeted quantitative mass spectrometry approach to profile changes of acetylation and methylation at multiple lysine residues on the N-terminal tail of histones H3 and H4. Our analyses identified a number of histone acetylation sites that are significantly affected by S. pombe HDAC mutations. We discovered that mutation of the Class I HDAC known as Clr6 causes a major increase in the abundance of triacetylated H4 molecules at K5, K8, and K12. A clr6-1 hypomorphic mutation also increased the abundance of multiple acetyl-lysines in histone H3. In addition, our study uncovered a few crosstalks between histone acetylation and methylation upon deletion of HDACs Hos2 and Clr3. We anticipate that the results from this study will greatly improve our current understanding of the mechanisms involved in HDAC-mediated gene regulation and heterochromatin assembly.
authors.affiliation: Department of Chemistry, ‡Institute for Research in Immunology and Cancer (IRIC), §Molecular Biology Programme, and ⊥Department of Pathology and Cell Biology, Université de Montréal , Montréal, Québec H3C 3J7, Canada.
authors.forename: Nebiyu
authors.initials: N
authors.lastname: Abshiru
authors.affiliation: Department of Chemistry, ‡Institute for Research in Immunology and Cancer (IRIC), §Molecular Biology Programme, and ⊥Department of Pathology and Cell Biology, Université de Montréal , Montréal, Québec H3C 3J7, Canada.
authors.forename: Roshan Elizabeth
authors.initials: RE
authors.lastname: Rajan
authors.affiliation: Department of Chemistry, ‡Institute for Research in Immunology and Cancer (IRIC), §Molecular Biology Programme, and ⊥Department of Pathology and Cell Biology, Université de Montréal , Montréal, Québec H3C 3J7, Canada.
authors.forename: Alain
authors.initials: A
authors.lastname: Verreault
authors.affiliation: Department of Chemistry, ‡Institute for Research in Immunology and Cancer (IRIC), §Molecular Biology Programme, and ⊥Department of Pathology and Cell Biology, Université de Montréal , Montréal, Québec H3C 3J7, Canada.
authors.forename: Pierre
authors.initials: P
authors.lastname: Thibault
mesh_terms: D000107:Acetylation; D042421:Histone Code; D006655:Histone Deacetylases / Q000235:genetics*; D006657:Histones / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D008745:Methylation; D009154:Mutation; D029702:Schizosaccharomyces pombe Proteins / Q000032:analysis / Q000378:metabolism*
chemical_list: D006657:Histones; D029702:Schizosaccharomyces pombe Proteins; D006655:Histone Deacetylases
keywords: chromatin; histone; histone deacetylase; mass spectrometry; post-translational modifications