title: Discovery of histone modification crosstalk networks by stable isotope labeling of amino acids in cell culture mass spectrometry (SILAC MS).
abstract: In this paper we describe an approach that combines stable isotope labeling of amino acids in cells culture, high mass accuracy liquid chromatography tandem mass spectrometry and a novel data analysis approach to accurately determine relative peptide post-translational modification levels. This paper describes the application of this approach to the discovery of novel histone modification crosstalk networks in Saccharomyces cerevisiae. Yeast histone mutants were generated to mimic the presence/absence of 44 well-known modifications on core histones H2A, H2B, H3, and H4. In each mutant strain the relative change in H3 K79 methylation and H3 K56 acetylation were determined using stable isotope labeling of amino acids in cells culture. This approach showed relative changes in H3 K79 methylation and H3 K56 acetylation that are consistent with known histone crosstalk networks. More importantly, this study revealed additional histone modification sites that affect H3 K79 methylation and H3 K56 acetylation.
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Guan
authors.forename: Neha
authors.initials: N
authors.lastname: Rastogi
authors.forename: Mark R
authors.initials: MR
authors.lastname: Parthun
authors.forename: Michael A
authors.initials: MA
authors.lastname: Freitas
mesh_terms: D000107:Acetylation; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D007553:Isotope Labeling; D008745:Methylation; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000596:Amino Acids; D006657:Histones; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins