Submitted to: FEBS Journal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/1/2005
Publication Date: 8/1/2005
Citation: Kobza, K., Kueh, A., Rueckert, B., Camporeale, G., Griffin, J.B., Sarath, G., Zempleni, J. 2005. K4, k9, and k18 are biotinylated in human histone h3. FEBS Journal 272(16):4249-4259. Interpretive Summary: Histones are DNA-binding proteins that are reversibly modified through covalent attachment of small molecules, such as phosphate or the vitamin biotin. Modification of histones by these small molecules causes chromatin remodeling, with resultant effects on critical cell processes. In this work, we have used small peptides and site-specific antibodies to study biotinylation of the histone - H3. We have found specific lysine residues that are modified by biotinylation, and demonstrated that these changes occur in the cell.
Technical Abstract: Histones are modified posttranslationally, 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. 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 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.