title: Preferential Phosphorylation on Old Histones during Early Mitosis in Human Cells.
abstract: How histone post-translational modifications (PTMs) are inherited through the cell cycle remains poorly understood. Canonical histones are made in the S phase of the cell cycle. Combining mass spectrometry-based technologies and stable isotope labeling by amino acids in cell culture, we question the distribution of multiple histone PTMs on old versus new histones in synchronized human cells. We show that histone PTMs can be grouped into three categories according to their distributions. Most lysine mono-methylation and acetylation PTMs are either symmetrically distributed on old and new histones or are enriched on new histones. In contrast, most di- and tri-methylation PTMs are enriched on old histones, suggesting that the inheritance of different PTMs is regulated distinctly. Intriguingly, old and new histones are distinct in their phosphorylation status during early mitosis in the following three human cell types: HeLa, 293T, and human foreskin fibroblast cells. The mitotic hallmark H3S10ph is predominantly associated with old H3 at early mitosis and becomes symmetric with the progression of mitosis. This same distribution was observed with other mitotic phosphorylation marks, including H3T3/T6ph, H3.1/2S28ph, and H1.4S26ph but not S28/S31ph on the H3 variant H3.3. Although H3S10ph often associates with the neighboring Lys-9 di- or tri-methylations, they are not required for the asymmetric distribution of Ser-10 phosphorylation on the same H3 tail. Inhibition of the kinase Aurora B does not change the distribution despite significant reduction of H3S10ph levels. However, K9me2 abundance on the new H3 is significantly reduced after Aurora B inhibition, suggesting a cross-talk between H3S10ph and H3K9me2.
authors.affiliation: From the Epigenetics Program, Department of Biochemistry and Biophysics, and.
authors.forename: Shu
authors.initials: S
authors.lastname: Lin
authors.affiliation: From the Epigenetics Program, Department of Biochemistry and Biophysics, and.
authors.forename: Zuo-Fei
authors.initials: ZF
authors.lastname: Yuan
authors.affiliation: From the Epigenetics Program, Department of Biochemistry and Biophysics, and.
authors.forename: Yumiao
authors.initials: Y
authors.lastname: Han
authors.affiliation: the Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104.
authors.forename: Dylan M
authors.initials: DM
authors.lastname: Marchione
authors.affiliation: From the Epigenetics Program, Department of Biochemistry and Biophysics, and bgarci@mail.med.upenn.edu.
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Garcia
mesh_terms: D064107:Aurora Kinase B / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D008938:Mitosis / Q000502:physiology*; D010766:Phosphorylation / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000502:physiology*
chemical_list: D006657:Histones; C576921:AURKB protein, human; D064107:Aurora Kinase B
keywords: chromatin; epigenetics; histone; mass spectrometry (MS); mitosis; post-translational modification (PTM); proteomics