The human decatenation checkpoint

Chromatid catenation is actively monitored in human cells, with progression from G2 to mitosis being inhibited when chromatids are insufficiently decatenated. Mitotic delay was quantified in normal and checkpoint-deficient human cells during treatment with ICRF-193, a topoisomerase II catalytic inhibitor that prevents chromatid decatenation without producing topoisomerase-associated DNA strand breaks. Ataxia telangiectasia (A-T) cells, defective in DNA damage checkpoints, showed normal mitotic delay when treated with ICRF-193. The mitotic delay in response to ICRF-193 was ablated in human fibroblasts expressing an ataxia telangiectasia mutated- and rad3-related (ATR) kinase-inactive ATR allele (ATRki). BRCA1-mutant HCC1937 cells also displayed a defect in ICRF-193-induced mitotic delay, which was corrected by expression of wild-type BRCA1. Phosphorylations of hCds1 or Chk1 and inhibition of Cdk1 kinase activity, which are elements of checkpoints associated with DNA damage or replication, did not occur during ICRF-193-induced mitotic delay. Over-expression of cyclin B1 containing a dominant nuclear localization signal, and inhibition of Crm1-mediated nuclear export, reversed ICRF-193-induced mitotic delay. In combination, these results imply that ATR and BRCA1 enforce the decatenation G2 checkpoint, which may act to exclude cyclin B1/Cdk1 complexes from the nucleus. Moreover, induction of ATRki produced a 10-fold increase in chromosomal aberrations, further emphasizing the vital role for ATR in genetic stability.

[1]  R. Paules,et al.  Inactivation of G2 checkpoint function and chromosomal destabilization are linked in human fibroblasts expressing human papillomavirus type 16 E6. , 1997, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[2]  E. Nigg,et al.  Cyclin‐dependent protein kinases: Key regulators of the eukaryotic cell cycle , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[3]  T. Andoh,et al.  Inhibition of intracellular topoisomerase II by antitumor bis(2,6-dioxopiperazine) derivatives: mode of cell growth inhibition distinct from that of cleavable complex-forming type inhibitors. , 1991, Cancer research.

[4]  J. Sarkaria,et al.  The radiosensitizing agent 7-hydroxystaurosporine (UCN-01) inhibits the DNA damage checkpoint kinase hChk1. , 2000, Cancer research.

[5]  J. Wang,et al.  Antitumor bisdioxopiperazines inhibit yeast DNA topoisomerase II by trapping the enzyme in the form of a closed protein clamp. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Pines.  Regulation of the G2 to M transition. , 1998, Results and problems in cell differentiation.

[7]  Bo Xu,et al.  Involvement of Brca1 in S-Phase and G2-Phase Checkpoints after Ionizing Irradiation , 2001, Molecular and Cellular Biology.

[8]  J. Sarkaria,et al.  Inhibition of ATM and ATR kinase activities by the radiosensitizing agent, caffeine. , 1999, Cancer research.

[9]  R. Paules,et al.  DNA damage and cell cycle checkpoints , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[10]  A. Fornace,et al.  The p53-dependent G1 cell cycle checkpoint pathway and ataxia-telangiectasia. , 1994, Cancer research.

[11]  Minoru Yoshida,et al.  CRM1 is responsible for intracellular transport mediated by the nuclear export signal , 1997, Nature.

[12]  D. Baltimore,et al.  ATR disruption leads to chromosomal fragmentation and early embryonic lethality. , 2000, Genes & development.

[13]  E. Nishida,et al.  Nuclear export of cyclin B1 and its possible role in the DNA damage‐induced G2 checkpoint , 1998, The EMBO journal.

[14]  C. Holm.  Coming undone: How to untangle a chromosome , 1994, Cell.

[15]  S. T. Kim,et al.  Substrate Specificities and Identification of Putative Substrates of ATM Kinase Family Members* , 1999, The Journal of Biological Chemistry.

[16]  Y. A. Minamishima,et al.  Aberrant cell cycle checkpoint function and early embryonic death in Chk1(-/-) mice. , 2000, Genes & development.

[17]  A. N. Meyer,et al.  Nuclear localization of cyclin B1 mediates its biological activity and is regulated by phosphorylation. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[18]  L. Hartwell,et al.  Cell cycle control and cancer. , 1994, Science.

[19]  A. Kumagai,et al.  Positive regulation of Wee1 by Chk1 and 14-3-3 proteins. , 2001, Molecular biology of the cell.

[20]  Edward A. Sausville,et al.  The Chk1 Protein Kinase and the Cdc25C Regulatory Pathways Are Targets of the Anticancer Agent UCN-01* , 2000, The Journal of Biological Chemistry.

[21]  Stephen J. Elledge,et al.  p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest , 1994, Cell.

[22]  H. Piwnica-Worms,et al.  ATR-Mediated Checkpoint Pathways Regulate Phosphorylation and Activation of Human Chk1 , 2001, Molecular and Cellular Biology.

[23]  S. Elledge,et al.  Functional interactions between BRCA1 and the checkpoint kinase ATR during genotoxic stress. , 2000, Genes & development.

[24]  D. Morgan,et al.  Nuclear Localization of Cyclin B1 Controls Mitotic Entry After DNA Damage , 1998, The Journal of cell biology.

[25]  J Chen,et al.  Ataxia telangiectasia-related protein is involved in the phosphorylation of BRCA1 following deoxyribonucleic acid damage. , 2000, Cancer research.

[26]  S. Elledge,et al.  Conservation of the Chk1 checkpoint pathway in mammals: linkage of DNA damage to Cdk regulation through Cdc25. , 1997, Science.

[27]  S. Elledge,et al.  Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint. , 2000, Genes & development.

[28]  M. Gatei,et al.  Chk1 complements the G2/M checkpoint defect and radiosensitivity of ataxia-telangiectasia cells , 1999, Oncogene.

[29]  E. Krebs,et al.  Mitogen-activated protein kinase kinase activity is required for the G(2)/M transition of the cell cycle in mammalian fibroblasts. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[30]  S. Schreiber,et al.  Overexpression of a kinase‐inactive ATR protein causes sensitivity to DNA‐damaging agents and defects in cell cycle checkpoints , 1998, The EMBO journal.

[31]  S. Elledge,et al.  Ataxia telangiectasia-mutated phosphorylates Chk2 in vivo and in vitro. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[32]  D. Scott,et al.  Cell death, chromosome damage and mitotic delay in normal human, ataxia telangiectasia and retinoblastoma fibroblasts after x-irradiation. , 1981, International journal of radiation biology and related studies in physics, chemistry, and medicine.

[33]  D. J. Clarke,et al.  A topoisomerase II-dependent G2 cycle checkpoint in mammalian cells , 1994, Nature.

[34]  K. Kuma,et al.  Isolation and characterization of the active cDNA of the human cell cycle gene (RCC1) involved in the regulation of onset of chromosome condensation. , 1987, Genes & development.

[35]  J. Moore,et al.  Control of cyclin B1 localization through regulated binding of the nuclear export factor CRM1. , 1998, Genes & development.

[36]  D. Livingston,et al.  Genetic analysis of BRCA1 function in a defined tumor cell line. , 1999, Molecular cell.

[37]  H. Piwnica-Worms,et al.  A human Cds1-related kinase that functions downstream of ATM protein in the cellular response to DNA damage. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[38]  C. Peng,et al.  Mitotic and G2 checkpoint control: regulation of 14-3-3 protein binding by phosphorylation of Cdc25C on serine-216. , 1997, Science.

[39]  X. Wang,et al.  Centrosome amplification and a defective G2-M cell cycle checkpoint induce genetic instability in BRCA1 exon 11 isoform-deficient cells. , 1999, Molecular cell.

[40]  H. Piwnica-Worms,et al.  Overproduction of Human Myt1 Kinase Induces a G2 Cell Cycle Delay by Interfering with the Intracellular Trafficking of Cdc2-Cyclin B1 Complexes , 1999, Molecular and Cellular Biology.

[41]  W. Kaufmann,et al.  DNA signals for G2 checkpoint response in diploid human fibroblasts. , 1998, Mutation research.

[42]  A. Kumagai,et al.  Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts. , 2000, Genes & development.

[43]  S. Elledge,et al.  BASC, a super complex of BRCA1-associated proteins involved in the recognition and repair of aberrant DNA structures. , 2000, Genes & development.

[44]  Y Taya,et al.  A role for ATR in the DNA damage-induced phosphorylation of p53. , 1999, Genes & development.

[45]  J. Raleigh,et al.  The G(2) DNA damage checkpoint targets both Wee1 and Cdc25. , 2000, Journal of cell science.

[46]  Amanda G Paulovich,et al.  When Checkpoints Fail , 1997, Cell.

[47]  J. Pines,et al.  MPF localization is controlled by nuclear export , 1998, The EMBO journal.

[48]  L. Hartwell,et al.  Checkpoints: controls that ensure the order of cell cycle events. , 1989, Science.

[49]  M. Roberge,et al.  Topoisomerase II inhibitors affect entry into mitosis and chromosome condensation in BHK cells. , 1996, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[50]  J. Pines,et al.  Translocation of cyclin B1 to the nucleus at prophase requires a phosphorylation-dependent nuclear import signal , 1999, Current Biology.

[51]  M. Gatei,et al.  Ataxia Telangiectasia Mutated (ATM) Kinase and ATM and Rad3 Related Kinase Mediate Phosphorylation of Brca1 at Distinct and Overlapping Sites , 2001, The Journal of Biological Chemistry.

[52]  David O. Morgan,et al.  Principles of CDK regulation , 1995, Nature.