Abstract
Checkpoint genes cause cell cycle arrest when DNA is damaged or DNA replication is blocked. Although a human homolog of Chk1 (hChk1) has recently been reported to be involved in the DNA damage checkpoint through phosphorylation of Cdc25A, B, and C, it is not known at which phase(s) of the cell cycle hChk1 functions and how hChk1 causes cell cycle arrest in response to DNA damage. In the present study, we demonstrate that in normal human fibroblasts (MJ90), hChk1 is expressed specifically at the S to M phase of the cell cycle at both the RNA and protein levels and that it is localized to the nucleus at this time. hChk1 activity, as determined by phosphorylation of Cdc25C, is readily detected at the S to M phase of the cell cycle, and DNA damage induced by UV or ionizing radiation does not enhance the expression of hChk1 or its activity. Furthermore, hChk1 exists in an active form at the S to M phase in fibroblasts derived from patients with ataxia telangiectasia (AT) which lack the functional AT mutated (ATM) gene product, suggesting that hChk1 expression is independent of functional ATM. Taken together with the findings that phosphorylation of Cdc25C on serine 216 is increased at the S to M phase, it is suggested that at this particular phase of the cell cycle, even in the absence of DNA damage, hChk1 phosphorylates Cdc25C on serine 216, which is considered to be a prerequisite for the G2/M checkpoint. Thus, hChk1 may play an important role in keeping Cdc25C prepared for responding to DNA damage by phosphorylating its serine residue at 216 during the S to M phase.
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References
Al-Khodairy F and Carr AM. . 1992 EMBO J. 11: 1343–1350.
Al-Khodairy F, Fotou E, Sheldrick KS, Griffiths DJ, Lehmann AR and Carr AM. . 1994 Mol. Biol. Cell. 5: 147–160.
Bentley NJ, Holtzman DA, Flaggs G, Keegan KS, DeMaggio A, Ford JC, Hoekstra M and Carr AM. . 1996 EMBO J. 15: 6641–6651.
Boddy MN, Furnari B, Mondesert O and Russell P. . 1998 Science 280: 909–912.
Carr AM. . 1996 Science 271: 314–315.
Cimprich KA, Shin TB, Keith CT and Schreiber SL. . 1996 Proc. Natl. Acad. Sci. USA 93: 2850–2855.
Cunliffe PN, Mann JR, Cameron AH, Roberts KD and Ward HWC. . 1975 Br. J. Radiobiol. 48: 374–376.
Elledge SJ. . 1996 Science 274: 1664–1672.
Enoch T, Carr AM and Nurse P. . 1992 Genes Dev. 6: 2035–2046.
Enoch T, Gould KL and Nurse P. . 1991 Cold Spring Harbor Symp. Quant. Biol. 56: 409–416.
Furnari B, Rhind N and Russell P. . 1997 Science 277: 1495–1497.
Flaggs G, Plug AW, Dunks KM, Mundt KE, Ford JC, Quiggle MR, Taylor EM, Westphal CH, Ashley T, Hoekstra MF and Carr AM. . 1997 Curr. Biol. 7: 977–986.
Gatti RA, Boder E, Vinters HV, Sparkes RS, Norman A and Lange K. . 1991 Medicin 70: 99–117.
Gotoff SP, Amirmokri E and Liebner EJ. . 1967 Am. J. Dis. Child. 114: 617–625.
Hartwell LH and Weinert TA. . 1989 Science 246: 629–634.
Hermeking H, Lengauer C, Polyak K, He T-C, Zhang L, Thiagalingam S, Kinzler KW and Vogelstein B. . 1997 Molec. Cell 1: 3–11.
Humphrey T and Enoch T. . 1995 Curr. Biol. 5: 376–378.
Kastan MB, Zhan O, El-Deiry WS, Carrier F, Jacks T, Walsh WV, Plunkett BS, Vogelstein B and Fornace AJ. . 1992 Cell 71: 587–597.
Khanna KK and Lavin MF. . 1993 Oncogene 8: 3307–3312.
Li JJ and Deshaies RJ. . 1993 Cell 74: 223–226.
Lyndall D and Weinert TA. . 1995 Science 270: 1488–1491.
Lindsay HD, Griffiths DJF, Edwards RJ, Christensen PU, Murray JM, Osman F, Walworth N and Carr AM. . 1998 Genes Dev. 12: 382–395.
Mockinnon PJ. . 1987 Hum. Genet. 75: 197–208.
Morgan JL, Holcomb TM and Morrissey RW. . 1968 Am. J. Dis. Child. 116: 557–558.
Murray AW. . 1992 Nature 359: 599–604.
Nakanishi M, Adami GR, Robetorye RS, Noda A, Venable SF, Dimitrov D, Pereira-Smith OM and Smith JR. . 1995 Proc. Natl. Acad. Sci. USA 92: 4532–4536.
Nurse P. . 1994 Cell 79: 547–550.
Peng C-Y, Graves PR, Thoma RS, Wu Z, Shaw AS and Piwnica-Worms H. . 1997 Science 277: 1501–1505.
Peng C-Y, Graves PR, Ogg S, Thoma RS, Byrnes III MJ, Wu Z, Stephenson MT and Piwnica-Worms H. . 1998 Cell Growth and Differ. 9: 197–208.
Rowley R, Subramani S and Young PG. . 1992 EMBO J. 11: 1335–1342.
Sanchez Y, Wong C, Thoma RS, Richman R, Wu Z, Piwnica-Worms H and Elledge SJ. . 1997 Science 277: 1497–1501.
Savitsky K, Bar-Shira A, Gilad S, Rotman G, Ziv Y, Vanagaite L, Tagle DA, Smith S, Uziel T, Sfez S, Ashkenazi M, Pecker I, Frydman M, Harnik R, Patanjali SR, Simmons A, Clines GA, Sartiel A, Gatti RA, Chessa L, Sanal O, Lavin MF, Jaspers NGJ, Tayler AMR, Arlett CF, Miki T, Weissman SM, Lovett M, Collins FS and Shiloh Y. . 1995 Science 268: 1749–1753.
Stewart E and Enoch T. . 1996 Curr. Opin. Cell Biol. 8: 781–787.
Taylor AMR, Harnden DG, Arlett CF, Harcourt AR, Lehmann AR, Stevens S and Bridges BA. . 1975 Nature 285: 427–429.
Walworth N, Daveyk S and Beach D. . 1993 Nature 363: 368–371.
Walworth NC and Bernards R. . 1996 Science 271: 353–356.
Watanabe N, Broome M and Hunter T. . 1995 EMBO J. 9: 1878–1891.
Weinert T. . 1997 Science 277: 1450–1451.
Acknowledgements
We thank Dr Takachika Azuma (Research Institute for Biological Sciences, Science University of Tokyo) for purifying an antibody against hChk1 protein. We also thank Mr Hiroshi Shimada, Drs Rikio Kato and Kengo Ito (Department of Radiology, National Chubu Hospital) for the irradiation of various cells. We appreciate Drs Mitsuhiro Yanagida and Fumiko Esashi (Graduate School of Science, Kyoto University) for discussion and providing valuable information before publication and the members of our cell cycle group in the Department of Geriatric Research, National Institute for Longevity Sciences, for discussions. This work was supported in part by a Grant-in-aid (to M Nakanishi) for Scientific Research on Priority Areas (No. 09273104) from the Ministry of Education, Science, Sports, and Culture of Japan, and a Health Sciences Research Grant for Research on Human Genome and Gene Therapy (No. H10-genome-001 to K Ikeda).
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Kaneko, Y., Watanabe, N., Morisaki, H. et al. Cell cycle-dependent and ATM-independent expression of human Chk1 kinase. Oncogene 18, 3673–3681 (1999). https://doi.org/10.1038/sj.onc.1202706
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DOI: https://doi.org/10.1038/sj.onc.1202706
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