Securin Is Required for Chromosomal Stability in Human Cells

Abnormalities of chromosome number are the most common genetic aberrations in cancer. The mechanisms regulating the fidelity of mitotic chromosome transmission in mammalian cells are therefore of great interest. Here we show that human cells without an hSecurin gene lose chromosomes at a high frequency. This loss was linked to abnormal anaphases during which cells underwent repetitive unsuccessful attempts to segregate their chromosomes. The abnormal mitoses were associated with biochemical defects in the activation of separin, the sister-separating protease, rendering it unable to cleave the cohesin subunit Scc1 efficiently. These results illuminate the function of mammalian securin and show that it is essential for the maintenance of euploidy.

[1]  Andrew W. Murray,et al.  Association of Spindle Assembly Checkpoint Component XMAD2 with Unattached Kinetochores , 1996, Science.

[2]  D. Burke,et al.  The spindle checkpoint: two transitions, two pathways. , 2000, Trends in cell biology.

[3]  G. Salvesen,et al.  Caspases - controlling intracellular signals by protease zymogen activation. , 2000, Biochimica et biophysica acta.

[4]  J. Herman,et al.  CpG methylation is maintained in human cancer cells lacking DNMT1 , 2000, Nature.

[5]  I. Sumara,et al.  Characterization of Vertebrate Cohesin Complexes and Their Regulation in Prophase , 2000, The Journal of cell biology.

[6]  David O. Morgan,et al.  Regulation of the APC and the exit from mitosis , 1999, Nature Cell Biology.

[7]  Bert Vogelstein,et al.  Mutations of mitotic checkpoint genes in human cancers , 1998, Nature.

[8]  Bert Vogelstein,et al.  Uncoupling of S phase and mitosis induced by anticancer agents in cells lacking p21 , 1996, Nature.

[9]  S. Melmed,et al.  Expression of pituitary-tumour transforming gene in colorectal tumours , 2000, The Lancet.

[10]  P. Duesberg,et al.  How aneuploidy may cause cancer and genetic instability. , 1999, Anticancer research.

[11]  K. Kinzler,et al.  Genetic instabilities in human cancers , 1998, Nature.

[12]  K. Kinzler,et al.  Genetic instability in colorectal cancers , 1997, Nature.

[13]  K Nasmyth,et al.  Splitting the chromosome: cutting the ties that bind sister chromatids. , 2000, Novartis Foundation symposium.

[14]  Y. Shiratori,et al.  Mutational Inactivation of Mitotic Checkpoint Genes, hsMAD2 and hBUB1, Is Rare in Sporadic Digestive Tract Cancers , 1999, Japanese journal of cancer research : Gann.

[15]  M. Yanagida,et al.  Fission yeast Cut1 and Cut2 are essential for sister chromatid separation, concentrate along the metaphase spindle and form large complexes. , 1996, The EMBO journal.

[16]  Marc W. Kirschner,et al.  How Proteolysis Drives the Cell Cycle , 1996, Science.

[17]  Stephen S. Taylor,et al.  The Human Homologue of Bub3 Is Required for Kinetochore Localization of Bub1 and a Mad3/Bub1-related Protein Kinase , 1998, The Journal of cell biology.

[18]  T. Kanda,et al.  Histone–GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells , 1998, Current Biology.

[19]  F. Ramos-Morales,et al.  hpttg, a human homologue of rat pttg, is overexpressed in hematopoietic neoplasms. Evidence for a transcriptional activation function of hPTTG , 1998, Oncogene.

[20]  C. Lehner,et al.  Separation of Sister Chromatids in Mitosis Requires the Drosophila pimples Product, a Protein Degraded after the Metaphase/Anaphase Transition , 1996, Cell.

[21]  A. Yoshimura,et al.  Somatic mutation of the hBUB1 mitotic checkpoint gene in primary lung cancer , 2000, Genes, chromosomes & cancer.

[22]  Kim Nasmyth,et al.  An ESP1/PDS1 Complex Regulates Loss of Sister Chromatid Cohesion at the Metaphase to Anaphase Transition in Yeast , 1998, Cell.

[23]  D. Ward,et al.  Karyotyping human chromosomes by combinatorial multi-fluor FISH , 1996, Nature Genetics.

[24]  Friedrich Lottspeich,et al.  Sister-chromatid separation at anaphase onset is promoted by cleavage of the cohesin subunit Scc1 , 1999, Nature.

[25]  L. Loeb,et al.  Mutator phenotype may be required for multistage carcinogenesis. , 1991, Cancer research.

[26]  Tim Hunt,et al.  Cut2 proteolysis required for sister-chromatid separation in fission yeast , 1996, Nature.

[27]  M. Kirschner,et al.  The checkpoint protein MAD2 and the mitotic regulator CDC20 form a ternary complex with the anaphase-promoting complex to control anaphase initiation. , 1998, Genes & development.

[28]  M. Yanagida,et al.  Cell cycle mechanisms of sister chromatid separation; Roles of Cut1/separin and Cut2/securin , 2000, Genes to cells : devoted to molecular & cellular mechanisms.

[29]  M. Kirschner,et al.  Anaphase initiation in Saccharomyces cerevisiae is controlled by the APC-dependent degradation of the anaphase inhibitor Pds1p. , 1996, Genes & development.

[30]  M. Glotzer.  Chromosome segregation: Samurai separation of Siamese sisters , 1999, Current Biology.

[31]  C. Lehner,et al.  Degradation of Drosophila PIM regulates sister chromatid separation during mitosis. , 2000, Genes & development.

[32]  T. Hirano,et al.  Identification of Xenopus SMC protein complexes required for sister chromatid cohesion. , 1998, Genes & development.

[33]  M. Kirschner,et al.  Identification of a vertebrate sister-chromatid separation inhibitor involved in transformation and tumorigenesis. , 1999, Science.

[34]  V. Guacci,et al.  Pds1p, an inhibitor of anaphase in budding yeast, plays a critical role in the APC and checkpoint pathway(s) , 1996, The Journal of cell biology.

[35]  P. Sorger,et al.  Retention of the BUB3 checkpoint protein on lagging chromosomes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[36]  W. Gerald,et al.  MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells , 2001, Nature.

[37]  P. Sorger,et al.  Chromosome Missegregation and Apoptosis in Mice Lacking the Mitotic Checkpoint Protein Mad2 , 2000, Cell.

[38]  Stephen S. Taylor,et al.  Kinetochore Localization of Murine Bub1 Is Required for Normal Mitotic Timing and Checkpoint Response to Spindle Damage , 1997, Cell.

[39]  R. Benezra,et al.  Identification of a Human Mitotic Checkpoint Gene: hsMAD2 , 1996, Science.

[40]  F. Ramos-Morales,et al.  hpttg is over-expressed in pituitary adenomas and other primary epithelial neoplasias , 1999, Oncogene.

[41]  K. Kinzler,et al.  Requirement for p53 and p21 to sustain G2 arrest after DNA damage. , 1998, Science.

[42]  M. Peinado,et al.  Cytogenetic characterization of two colon cell lines by using conventional G-banding, comparative genomic hybridization, and whole chromosome painting. , 2000, Cancer genetics and cytogenetics.

[43]  J. Peters,et al.  Subunits and substrates of the anaphase-promoting complex. , 1999, Experimental cell research.

[44]  B. Ponder,et al.  Mitotic checkpoint inactivation fosters transformation in cells lacking the breast cancer susceptibility gene, Brca2. , 1999, Molecular cell.

[45]  V. Guacci,et al.  Pds1p is required for faithful execution of anaphase in the yeast, Saccharomyces cerevisiae , 1996, The Journal of cell biology.

[46]  Kim Nasmyth,et al.  Cleavage of Cohesin by the CD Clan Protease Separin Triggers Anaphase in Yeast , 2000, Cell.

[47]  C Lengauer,et al.  Genetic instability and darwinian selection in tumours. , 1999, Trends in cell biology.