Phosphorylation of distinct regions of f1 histone. Relationship to the cell cycle.
The phosphorylation of different amino acids in distinct regions of f1 histone was studied in highly synchronized Chinese hamster cell populations (line CHO). The purified, 32P-labeled f1 histone was bisected into NH2-terminal and COOH-terminal fragments with N-bromosuccinimide. Tryptic phosphopeptides from these fragments were resolved using sequential high voltage electrophoretic steps on paper. No phosphorylation was observed in early G1-arrested cells. Interphase phosphorylation began in late G1 in the COOH-terminal portion of the molecule on serine. This event continued throughout S phase and persisted into mitosis. However, in mitosis additional phosphorylation was observed in the COOH-terminal portion of the molecule on threonine, and for the only time in the CHO cell cycle the NH2-terminal portion of the molecule was also phosphorylated on both serine and threonine. The peptide studies thus predicted that a minimum of four sites (two serine and two threonine) were phosphorylated in the f1 histone of mitotic CHO cells. This was confirmed using electrophoresis in long polyacrylamide gels.
D000596:Amino Acids / Q000032:analysis; D002455:Cell Division; D002460:Cell Line; D006657:Histones / Q000378:metabolism*; D046911:Macromolecular Substances; D008938:Mitosis; D008970:Molecular Weight; D010085:Oxidative Phosphorylation; D010446:Peptide Fragments / Q000032:analysis; D006656:Protamine Kinase / Q000378:metabolism*; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D013997:Time Factors; D014357:Trypsin
D000596:Amino Acids; D006657:Histones; D046911:Macromolecular Substances; D010446:Peptide Fragments; D011494:Protein Kinases; D006656:Protamine Kinase; D014357:Trypsin