title: K4, K9 and K18 in human histone H3 are targets for biotinylation by biotinidase.
abstract: Histones are modified post-translationally, e.g. by methylation of lysine and arginine residues, and by phosphorylation of serine residues. These modifications regulate processes such as gene expression, DNA repair, and mitosis and meiosis. Recently, evidence has been provided that histones are also modified by covalent binding of the vitamin biotin. The aims of this study were to identify biotinylation sites in histone H3, and to investigate the crosstalk among histone biotinylation, methylation and phosphorylation. Synthetic peptides based on the sequence of human histone H3 were used as substrates for enzymatic biotinylation by biotinidase; biotin in peptides was probed using streptavidin peroxidase. These studies provided evidence that K4, K9 and K18 in histone H3 are good targets for biotinylation; K14 and K23 are relatively poor targets. Antibodies were generated to histone H3, biotinylated either at K4, K9 or K18. These antibodies localized to nuclei in human placental cells in immunocytochemistry and immunoblotting experiments, suggesting that lysines in histone H3 are biotinylated in vivo. Dimethylation of R2, R8 and R17 increased biotinylation of K4, K9 and K18, respectively, by biotinidase; phosphorylation of S10 abolished biotinylation of K9. These observations are consistent with crosstalk between biotinylation of histones and other known modifications of histones. We speculate that this crosstalk provides a link to known roles for biotin in gene expression and cell proliferation.
authors.affiliation: Department of Nutrition and Health Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583-0806, USA.
authors.forename: Keyna
authors.initials: K
authors.lastname: Kobza
authors.forename: Gabriela
authors.initials: G
authors.lastname: Camporeale
authors.forename: Brian
authors.initials: B
authors.lastname: Rueckert
authors.forename: Alice
authors.initials: A
authors.lastname: Kueh
authors.forename: Jacob B
authors.initials: JB
authors.lastname: Griffin
authors.forename: Gautam
authors.initials: G
authors.lastname: Sarath
authors.forename: Janos
authors.initials: J
authors.lastname: Zempleni
mesh_terms: D000595:Amino Acid Sequence; D001710:Biotin / Q000378:metabolism*; D043523:Biotinidase / Q000378:metabolism*; D015153:Blotting, Western; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008745:Methylation; D008969:Molecular Sequence Data; D010766:Phosphorylation
chemical_list: D006657:Histones; D001710:Biotin; D043523:Biotinidase