title: In vivo residue-specific histone methylation dynamics.
abstract: Methylation of specific histone residues is capable of both gene activation and silencing. Despite vast work on the function of methylation, most studies either present a static snapshot of methylation or fail to assign kinetic information to specific residues. Using liquid chromatography-tandem mass spectrometry on a high-resolution mass spectrometer and heavy methyl-SILAC labeling, we studied site-specific histone lysine and arginine methylation dynamics. The detection of labeled intermediates within a methylation state revealed that mono-, di-, and trimethylated residues generally have progressively slower rates of formation. Furthermore, methylations associated with active genes have faster rates than methylations associated with silent genes. Finally, the presence of both an active and silencing mark on the same peptide results in a slower rate of methylation than the presence of either mark alone. Here we show that quantitative proteomic approaches such as this can determine the dynamics of multiple methylated residues, an understudied portion of histone biology.
authors: Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
authors: Barry M
authors: BM
authors: Zee
authors: Rebecca S
authors: RS
authors: Levin
authors: Bo
authors: B
authors: Xu
authors: Gary
authors: G
authors: LeRoy
authors: Ned S
authors: NS
authors: Wingreen
authors: Benjamin A
authors: BA
authors: Garcia
mesh_terms: D001120:Arginine / Q000737:chemistry; D001671:Biochemistry / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D020868:Gene Silencing; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008745:Methylation; D010455:Peptides / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods
chemical_list: D006657:Histones; D010455:Peptides; D001120:Arginine; D008239:Lysine