Browse > Article
http://dx.doi.org/10.5352/JLS.2006.16.1.012

Impaired Spindle Checkpoint Response of Brca1-deficient Mouse Embryonic Fibroblasts (MEFs) to Nocodazole Treatment  

Kim Myoung-Ae (Medical Research Center for Cancer Molecular Therapy, College of Medicine, Dong-A University, Department of Biochemistry, College of Medicine, Dong-A University)
Kim Hyunju (Medical Research Center for Cancer Molecular Therapy, College of Medicine, Dong-A University, Department of Biochemistry, College of Medicine, Dong-A University)
Yun Jeanho (Medical Research Center for Cancer Molecular Therapy, College of Medicine, Dong-A University, Department of Biochemistry, College of Medicine, Dong-A University)
Publication Information
Journal of Life Science / v.16, no.1, 2006 , pp. 12-16 More about this Journal
Abstract
Genetic alternation of Brca1 predispose of breast and ovarian cancer. Brca1 plays critical role in cell cycle regulation following DNA damage. Previous studies revealed that Brca1 plays an important role in S phase and G2/M checkpoint regulation. However, whether Brca1 involves in spindle checkpoint is unclear. In this study, the role of Brca1 in cell cycle response following nocodazole, which is a reagent that depolymerizes microtubules and activates the spindle checkpoint, has been examined using wild type $p53^{-/-}\;and\;p53^{-/-}Brca1^{-/-}$ mouse embryonic fibroblasts (MEFs). While wild type and Brca1-proficient MEFs showed an acute mitotic arrest, Brca1-deficient MEFs failed to arrest at mitotic phase in response to nocodazole treatment. In double-thymidine block and nocodazole treatment experiment, a portion of $p53^{-/-}\;Brca1^{-/-}$ MEFs were clearly by-passed nocodazole induced mitotic arrest. Consistent with this, in morphologic analysis, $p53^{-/-}\;Brca1^{-/-}$ MEFs showed growing cell morphology after nocodazole treatment. Taken together, these results suggest that Brca1 protein is an important component for normal induction of spindlecheckpoint and impairment of Brca1 function could induce dysregulation of mitotic cell cycle that ultimately results in genomic instability.
Keywords
Brca1; Spindle checkpoint; nocodazole; cell cycle;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Gasco, M., Yulug, I. G., and Crook, T. 2003. TP53 mutations in familial breast cancer: functional aspects. Hum. Mutat. 21, 301-306   DOI   ScienceOn
2 Sablina AA, Agapova LS, ChumakovPM, and Kopnin BP. 1999. p53 does not control the spindle assembly cell cycle checkpoint but mediates G1 arrest in response to disruption of microtubule system. Cell. Biol. Int. 23, 323-334   DOI   ScienceOn
3 Nigg, E. A. 2001. Mitotic kinases as regulators of cell division and its checkpoints. Nat. Rev. Mol. Cell. Biol. 2, 21-32   DOI   ScienceOn
4 Vogel C, Kienitz A, Hofmann I, Muller R, and Bastians H. 2004. Crosstalk of the mitotic spindle assembly checkpoint with p53 to prevent polyploidy. Oncogene 23, 6845-6853   DOI   ScienceOn
5 Zheng, L., Li, S., Boyer, T. G., and Lee, W. H. 2000. Lessons learned from BRCA1 and BRCA2. Oncogene 19, 6159-6175   DOI
6 Xu, X., Qiao, W., Linke, S. P., Cao, L., Li, W. M., Furth, P. A., Harris, C. C., and Deng, C. X. 2001. Genetic interactions between tumor suppressors Brca1 and p53 in apoptosis, cell cycle and tumorigenesis. Nat. Genet. 28, 266-271   DOI   ScienceOn
7 Yun, J. Involvement of Brca1 in DNA interstrand crosslink repair through homologous recombination-independent process. J. Life Sci. 15, 542-547   DOI   ScienceOn
8 Yun, J., Zhong, Q., Kwak, J. Y., and Lee, W. H. 2005. Hypersensitivity of Brca1-deficient MEF to the DNA interstrand crosslinking agent mitomycin C is associated with defect in homologous recombination repair and aberrant S-phase arrest. Oncogene 24, 4009-4016   DOI   ScienceOn
9 Zheng, L., Pan, H., Li, S., Flesken-Nikitin, A., Chen, P. L., Boyer, T. G., and Lee, W. H. 2000. Sequence-specific transcriptional corepressor function for BRCA1 through a novel zinc finger protein, ZBRK1. Mol. Cell. 6, 757-768   DOI   ScienceOn
10 Hartwell, L. 1992. Defects in a cell cycle checkpoint may be responsible for the genomic instability of cancer cells. Cell 71, 543-546   DOI   ScienceOn
11 Jallepalli, P. V., and Lengauer, C. 2001. Chromosome segregation and cancer: cutting through the mystery. Nat. Rev. Cancer 1, 109-117   DOI   ScienceOn
12 Khan, S. H., and Wahl, G. M. 1998. p53 and pRb prevent rereplication in response to microtubule inhibitors by mediating a reversible G1 arrest. Cancer Res. 58, 396-401
13 Lanni, J. S., and Jacks, T. 1998. Characterization of the p53-dependent postmitotic checkpoint following spindle disruption. Mol. Cell. Biol. 18, 1055-1064   DOI
14 Lengauer, C., Kinzler, K. W., and Vogelstein, B. 1998. Genetic instabilities in human cancers. Nature 396, 643-649   DOI   ScienceOn
15 Ruffner, H., and Verma, I. M. 1997. BRCA1 is a cell cycle-regulated nuclear phosphoprotein. Proc. Natl. Acad. Sci. U. S. A. 94, 7138-7143
16 Brodie, S. G., Xu, X., Qiao, W., Li, W. M., Cao, L., and Deng, C. X. 2001. Multiple genetic changes are associated with mammary tumorigenesis in Brca1 conditional knockout mice. Oncogene 20, 7514-7523   DOI
17 Vaughn, J. P., Davis, P. L., Jarboe, M. D., Huper, G., Evans, A. C., Wiseman, R. W., Berchuck, A., Iglehart, J. D., Futreal, P. A., and Marks, J. R. 1996. BRCA1 expression isinduced before DNA synthesis in both normal and tumor-derived breast cells. Cell Growth Differ. 7, 711-715
18 Weaver, Z., Montagna, C., Xu, X., Howard, T., Gadina, M., Brodie, S. G., Deng, C. X., and Ried, T. 2002. Mammary tumors in mice conditionally mutant for Brca1 exhibit gross genomic instability and centrosome amplification yet display a recurring distribution of genomic imbalances that is similar to human breast cancer. Oncogene 21, 5097-5107   DOI   ScienceOn
19 Wang, R. H., Yu, H., and Deng, C. X. 2004. A requirement for breast-cancer-associated gene 1 (BRCA1) in the spindle checkpoint. Proc. Natl. Acad. Sci. U. S. A. 101, 17108-17113
20 Ting, N. S., and Lee, W. H. 2004. The DNA double-strand break response pathway: becoming more BRCAish than ever. DNA Repair (Amst), 3, 935-944   DOI   ScienceOn
21 Cross, S. M., Sanchez, C. A., Morgan, C. A., Schimke, M. K., Ramel, S., Idzerda, R. L., Raskind, W. H., and Reid, B. J. 1995. A p53-dependent mouse spindle checkpoint. Science 267, 1353-1356   DOI   ScienceOn
22 Chen, Y., Farmer, A. A., Chen, C. F., Jones, D. C., Chen, P. L., and Lee, W. H. 1996. BRCA1 is a 220-kDa nuclear phosphoprotein that is expressed and phosphorylated in a cell cycle-dependent manner. Cancer Res. 56, 3168-3172
23 Deng, C. X., and Brodie, S. G. 2000. Roles of BRCA1 and its interacting proteins. Bioassays 22, 728-737   DOI   ScienceOn
24 Di Leonardo, A., Khan, S. H., Linke, S. P., Greco, V., Seidita, G., and Wahl, G. M. 1997. DNA rereplication in the presence of mitotic spindle inhibitors in human and mouse fibroblasts lacking either p53 or pRb function. Cancer Res. 57, 1013-1019
25 Xu, X., Weaver, Z., Linke, S. P., Li, C., Gotay, J., Wang, X. W., Harris, C. C., Ried, T., and Deng, C. X. 1999. Centrosome amplification and a defective G2-M cell cycle checkpoint induce genetic instability in BRCA1 exon 11 isoform-deficient cells. Mol. Cell. 3, 389-395   DOI   ScienceOn