title: Modifications of human histone H3 variants during mitosis.
abstract: Phosphorylation of histone H3 is a hallmark event in mitosis and is associated with chromosome condensation. Here, we use a combination of immobilized metal affinity chromatography and tandem mass spectrometry to characterize post-translational modifications associated with phosphorylation on the N-terminal tails of histone H3 variants purified from mitotically arrested HeLa cells. Modifications observed in vivo on lysine residues adjacent to phosphorylated Ser and Thr provide support for the existence of the "methyl/phos", binary-switch hypothesis [Fischle, W., Wang, Y., and Allis, C. D. (2003) Nature 425, 475-479]. ELISA with antibodies selective for H3 at Ser10, Ser28, and Thr3 show reduced activity when adjacent Lys residues are modified. When used together, mass spectrometry and immunoassay methods provide a powerful approach for elucidation of the histone code and identification of histone post-translational modifications that occur during mitosis and other specific cellular events.
authors.affiliation: Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, USA.
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Garcia
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Barber
authors.forename: Sandra B
authors.initials: SB
authors.lastname: Hake
authors.forename: Celeste
authors.initials: C
authors.lastname: Ptak
authors.forename: Fiona B
authors.initials: FB
authors.lastname: Turner
authors.forename: Scott A
authors.initials: SA
authors.lastname: Busby
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Shabanowitz
authors.forename: Richard G
authors.initials: RG
authors.lastname: Moran
authors.forename: C David
authors.initials: CD
authors.lastname: Allis
authors.forename: Donald F
authors.initials: DF
authors.lastname: Hunt
mesh_terms: D002846:Chromatography, Affinity; D014644:Genetic Variation; D006367:HeLa Cells; D006657:Histones / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D013058:Mass Spectrometry; D008938:Mitosis; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational
chemical_list: D006657:Histones