title: Mapping post-translational modifications of the histone variant MacroH2A1 using tandem mass spectrometry.
abstract: Post-translational histone modifications modulate chromatin-templated processes and therefore affect cellular proliferation, growth, and development. Although post-translational modifications on the core histones have been under intense investigation for several years, the modifications on variant histones are poorly understood. We used tandem mass spectrometry to identify covalent modifications on a histone H2A variant, macroH2A1.2. MacroH2A1.2 can be monoubiquitinated; however, the site of monoubiquitination has not been documented. In this study we used green fluorescent protein-tagged macroH2A1.2 to determine that Lys(115) is a site of ubiquitination. In addition, we found that this variant H2A is methylated on the epsilon amino group of lysine residues Lys(17), Lys(122), and Lys(238) and phosphorylated on Thr(128). Three of these modifications were also found to be present in the endogenous protein by mass spectrometric analysis. These results provide the first direct evidence that multiple post-translational modifications are imposed on macroH2A1.2, suggesting that, like canonical H2A, this variant H2A is subject to regulation by combinatorial use of covalent modifications.
authors.affiliation: Mass Spectrometry Facility and Department of Pharmaceutical Chemistry, University of California, San Francisco, 94143, USA.
authors.forename: Feixia
authors.initials: F
authors.lastname: Chu
authors.forename: Dmitri A
authors.initials: DA
authors.lastname: Nusinow
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Chalkley
authors.forename: Kathrin
authors.initials: K
authors.lastname: Plath
authors.forename: Barbara
authors.initials: B
authors.lastname: Panning
authors.forename: Alma L
authors.initials: AL
authors.lastname: Burlingame
mesh_terms: D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D047468:Immunoprecipitation; D008239:Lysine / Q000737:chemistry; D008745:Methylation; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D021241:Spectrometry, Mass, Electrospray Ionization; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D006657:Histones; D025801:Ubiquitin; C120756:macroH2A histone; D008239:Lysine