title: Chromatin proteins captured by ChIP-mass spectrometry are linked to dosage compensation in Drosophila.
abstract: X-chromosome dosage compensation by the MSL (male-specific lethal) complex is required in Drosophila melanogaster to increase gene expression from the single male X to equal that of both female X chromosomes. Instead of focusing solely on protein complexes released from DNA, here we used chromatin-interacting protein MS (ChIP-MS) to identify MSL interactions on cross-linked chromatin. We identified MSL-enriched histone modifications, including histone H4 Lys16 acetylation and histone H3 Lys36 methylation, and CG4747, a putative Lys36-trimethylated histone H3 (H3K36me3)-binding protein. CG4747 is associated with the bodies of active genes, coincident with H3K36me3, and is mislocalized in the Set2 mutant lacking H3K36me3. CG4747 loss of function in vivo results in partial mislocalization of the MSL complex to autosomes, and RNA interference experiments confirm that CG4747 and Set2 function together to facilitate targeting of the MSL complex to active genes, validating the ChIP-MS approach.
authors.affiliation: Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts, USA.
authors.forename: Charlotte I
authors.initials: CI
authors.lastname: Wang
authors.forename: Artyom A
authors.initials: AA
authors.lastname: Alekseyenko
authors.forename: Gary
authors.initials: G
authors.lastname: LeRoy
authors.forename: Andrew E H
authors.initials: AE
authors.lastname: Elia
authors.forename: Andrey A
authors.initials: AA
authors.lastname: Gorchakov
authors.forename: Laura-Mae P
authors.initials: LM
authors.lastname: Britton
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Elledge
authors.forename: Peter V
authors.initials: PV
authors.lastname: Kharchenko
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Garcia
authors.forename: Mitzi I
authors.initials: MI
authors.lastname: Kuroda
mesh_terms: D000107:Acetylation; D000818:Animals; D030801:Animals, Genetically Modified; D015153:Blotting, Western; D047369:Chromatin Immunoprecipitation; D004303:Dosage Compensation, Genetic / Q000235:genetics*; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics*; D005260:Female; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008745:Methylation; D009687:Nuclear Proteins; D010088:Oxidoreductases / Q000378:metabolism*; D034622:RNA Interference
chemical_list: D029721:Drosophila Proteins; D006657:Histones; D009687:Nuclear Proteins; D010088:Oxidoreductases; C579351:NDF protein, Drosophila; D011495:Histone-Lysine N-Methyltransferase; C521696:Set2 protein, Drosophila