Cell Cycle Phase-specific Phosphorylation of Human Topoisomerase IIα EVIDENCE OF A ROLE FOR PROTEIN KINASE C
作者： Nicholas J. Wells , Andrew M. Fry , Fulvio Guano , Chris Norbury , Ian D. Hickson
关键词:
摘要: Type II topoisomerases are essential for faithful cell division in all organisms. In human cells, the α isozyme of topoisomerase has been implicated catalyzing mitotic chromosome segregation via its action as a DNA unlinking enzyme. Here, we have shown that enzymatic activity IIα protein purified from HeLa nuclei was strongly enhanced following phosphorylation by kinase C. We investigated possibility this is involved cycle phase-specific cells. Two-dimensional tryptic phosphopeptide mapping revealed immunoprecipitated metabolically labeled cells differentially phosphorylated during G2/M phases cycle. To identify sites and kinase(s) responsible modification, oligohistidine-tagged recombinant domains were overexpressed Escherichia coli affinity chromatography. Phosphorylation short fragment N-terminal ATPase domain C vitro generated two phosphopeptides co-migrated with prominent cell-derived protein. Site-directed mutagenesis studies indicated serine 29 both these phosphopeptides. Our results implicate phase-dependent modulation
sci-hub.se 参考文章(60)
M.E. Cardenas, Q. Dang, C.V. Glover, S.M. Gasser, Casein kinase II phosphorylates the eukaryote-specific C-terminal domain of topoisomerase II in vivo. The EMBO Journal. ,vol. 11, pp. 1785- 1796 ,(1992) , 10.1002/J.1460-2075.1992.TB05230.X
T. Uemura, M. Yanagida, Isolation of type I and II DNA topoisomerase mutants from fission yeast: single and double mutants show different phenotypes in cell growth and chromatin organization. The EMBO Journal. ,vol. 3, pp. 1737- 1744 ,(1984) , 10.1002/J.1460-2075.1984.TB02040.X
Serine 1524 is a major site of phosphorylation on human topoisomerase II alpha protein in vivo and is a substrate for casein kinase II in vitro. Journal of Biological Chemistry. ,vol. 269, pp. 29746- 29751 ,(1994) , 10.1016/S0021-9258(18)43944-0
D J Kroll, T C Rowe, Phosphorylation of DNA topoisomerase II in a human tumor cell line. Journal of Biological Chemistry. ,vol. 266, pp. 7957- 7961 ,(1991) , 10.1016/S0021-9258(20)89542-8
K Kimura, A Kikuchi, T Enomoto, M Saijo, M Ui, N Nozaki, Identification of the nature of modification that causes the shift of DNA topoisomerase II beta to apparent higher molecular weight forms in the M phase. Journal of Biological Chemistry. ,vol. 269, pp. 24523- 24526 ,(1994) , 10.1016/S0021-9258(17)31419-9
D G Campbell, D G Hardie, P R Vulliet, Identification of four phosphorylation sites in the N-terminal region of tyrosine hydroxylase. Journal of Biological Chemistry. ,vol. 261, pp. 10489- 10492 ,(1986) , 10.1016/S0021-9258(18)67410-1
N.R. Murray, G.P. Baumgardner, D.J. Burns, A.P. Fields, Protein kinase C isotypes in human erythroleukemia (K562) cell proliferation and differentiation. Evidence that beta II protein kinase C is required for proliferation. Journal of Biological Chemistry. ,vol. 268, pp. 15847- 15853 ,(1993) , 10.1016/S0021-9258(18)82331-6
P Ackerman, C V Glover, N Osheroff, Phosphorylation of DNA topoisomerase II in vivo and in total homogenates of Drosophila Kc cells. The role of casein kinase II. Journal of Biological Chemistry. ,vol. 263, pp. 12653- 12660 ,(1988) , 10.1016/S0021-9258(18)37803-7
P M Watt, I D Hickson, Structure and function of type II DNA topoisomerases. Biochemical Journal. ,vol. 303, pp. 681- 695 ,(1994) , 10.1042/BJ3030681
FH Drake, JP Zimmerman, FL McCabe, HF Bartus, SR Per, DM Sullivan, WE Ross, MR Mattern, RK Johnson, ST Crooke, Purification of topoisomerase II from amsacrine-resistant P388 leukemia cells. Evidence for two forms of the enzyme. Journal of Biological Chemistry. ,vol. 262, pp. 16739- 16747 ,(1987) , 10.1016/S0021-9258(18)49317-9