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Knock-down of human MutY homolog (hMYH) decreases phosphorylation of checkpoint kinase 1 (Chk1) induced by hydroxyurea and UV treatment

  • Hahm, Soo-Hyun (Department of Advanced Technology Fusion, Konkuk University) ;
  • Park, Jong-Hwa (Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University) ;
  • Ko, Sung-Il (Department of Advanced Technology Fusion, Konkuk University) ;
  • Lee, You-Ri (Department of Advanced Technology Fusion, Konkuk University) ;
  • Chung, In-Sik (Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University) ;
  • Chung, Ji-Hyung (Yonsei Integrative Research Institute for Cerebral & Cardiovascular Diseases (YIRIC), Yonsei University Health System) ;
  • Kang, Lin-Woo (Department of Advanced Technology Fusion, Konkuk University) ;
  • Han, Ye-Sun (Department of Advanced Technology Fusion, Konkuk University)
  • Received : 2010.12.18
  • Accepted : 2011.03.15
  • Published : 2011.05.31

Abstract

The effect of human MutY homolog (hMYH) on the activation of checkpoint proteins in response to hydroxyurea (HU) and ultraviolet (UV) treatment was investigated in hMYH-disrupted HEK293 cells. hMYH-disrupted cells decreased the phosphorylation of Chk1 upon HU or UV treatment and increased the phosphorylation of Cdk2 and the amount of Cdc25A, but not Cdc25C. In siMYH-transfected cells, the increased rate of phosphorylated Chk1 upon HU or UV treatment was lower than that in siGFP-transfected cells, meaning that hMYH was involved in the activation mechanism of Chk1 upon DNA damage. The phosphorylation of ataxia telangiectasia and Rad3-related protein (ATR) upon HU or UV treatment was decreased in hMYH-disrupted HEK293 and HaCaT cells. Co-immunoprecipitation experiments showed that hMYH was immunoprecipitated by anti-ATR. These results suggest that hMYH may interact with ATR and function as a mediator of Chk1 phosphorylation in response to DNA damage.

Keywords

References

  1. Jeggo, P. A. and Lobrich, M. (2006) Contribution of DNA repair and cell cycle checkpoint arrest to the maintenance of genomic stability. DNA Repair 5, 1192-1198. https://doi.org/10.1016/j.dnarep.2006.05.011
  2. Ohtsubo, T., Nishioka, K., Imaiso, Y., Iwai, S., Shimokawa, H., Oda, H., Fujiwara, T. and Nakabeppu, Y. (2000) Identification of human MutY homolog (hMYH) as a repair enzyme for 2-hydroxyadenine in DNA and detection of multiple forms of hMYH located in nuclei and mitochondria. Nucleic Acid Res. 28, 1355-1364. https://doi.org/10.1093/nar/28.6.1355
  3. Shinmura, K., Kasai, H., Sasaki, A., Sugimura, H. and Yokota, J. (1997) 8-hydroxyguanine (7,8-dihydro-8-oxoguanine) DNA glycosylase and AP lyase activities of hOGG1 protein and their substrate specificity. Mutat. Res. 385, 75-82.
  4. Sakumi, K., Furuichi, M., Tsuzuki, T., Kakuma, T., Kawabata, S. L., Maki, H. and Sekiguchi, M. (1993) Cloning and expression of cDNA for a human enzyme that hydrolyzes 8-oxo-dGTP, a mutagenic substrate for DNA synthesis. J. Biol. Chem. 268, 23524-23530.
  5. Xie, Y., Yang, H., Cunanan, C., Okamoto, K., Shibata, D., Pan, J., Barnes, D. E., Lindahl, T., McIlhatton, M., Fishel, R. and Miller, J. H. (2004) Deficiencies in mouse Myh and Ogg1 result in tumor predisposition and G to T mutations in codon 12 of the K-ras oncogene in lung tumors. Cancer Res. 64, 3096-3102. https://doi.org/10.1158/0008-5472.CAN-03-3834
  6. Sakamoto, K., Tominaga, Y., Yamauchi, K., Nakatsu, Y., Sakumi, K., Yoshiyama, K., Egashira, A., Kura, S., Yao, T., Tsuneyoshi, M., Maki, H., Nakabeppu, Y. and Tsuzuki, T. (2007) MUTYH-null mice are susceptible to spontaneous and oxidative stress induced intestinal tumorigenesis. Cancer Res. 67, 6599-6604. https://doi.org/10.1158/0008-5472.CAN-06-4802
  7. Xie, Y., Yang, H., Miller, J. H., Shih, D. M., Hicks, G. G., Xie, J. and Shiu, R. P. (2008) Cells deficient in oxidative DNA damage repair genes Myh and Ogg1 are sensitive to oxidants with increased G2/M arrest and multinucleation. Carcinogenesis 29, 722-728. https://doi.org/10.1093/carcin/bgn033
  8. Shi, G., Chang, D. Y., Cheng, C. C., Venclovas, C. and Lu, A. L. (2006) Physical and functional interactions between MutY glycosylase homologue (MYH) and checkpoint proteins Rad9-Rad1-Hus1. Biochem. J. 400, 53-62. https://doi.org/10.1042/BJ20060774
  9. Zhao, H. and Piwnica-Worms, H. (2001) ATR-mediated checkpoint pathways regulate phosphorylation and activation of human Chk1. Mol. Cell. Biol. 21, 4129-4139. https://doi.org/10.1128/MCB.21.13.4129-4139.2001
  10. Muller, B., Blackburn, J., Feijon, C., Zhao, X. and Smythe, C. (2007) DNA-activated protein kinase functions in a newly observed S phase checkpoint links histone mRNA abundance with DNA replication. J. Cell Biol. 31, 1385-1398.
  11. Walworth, N. C. (2000) Cell-cycle checkpoint kinase: checking in on cell cycle. Curr. Opin. Cell Biol. 12, 697-704. https://doi.org/10.1016/S0955-0674(00)00154-X
  12. Bartek, J. and Lukas, J. (2003) Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell 3, 421-429. https://doi.org/10.1016/S1535-6108(03)00110-7
  13. Donzelli, M. and Draetta, G. F. (2003) Regulating mammalian checkpoint through Cdc25 inactivation. EMBO Rep. 4, 671-677. https://doi.org/10.1038/sj.embor.embor887
  14. Donzelli, M., Squatrito, M., Ganoth, D., Hershko, A., Pagano, M. and Draetto, G. F. (2002) Dual mode of degradation of Cdc25 A phosphatase. EMBO J. 21, 2875-4884.
  15. Boldogh, I., Milligan, D., Lee, M. S., Bassett, H., Lloyd, R. S. and McCullough, A. K. (2001) hMYH cell cycle-dependent expression, subcellular localization and association with replication foci: evidence suggesting replication- coupled repair of adenine:8-oxoguanine mispairs. Nucleic Acid Res. 29, 2802-2809. https://doi.org/10.1093/nar/29.13.2802
  16. Ling, H., Wen, L., Ji, X. X., Tang, Y. L., He, J., Tan, H., Xia, H., Zhou, J. G. and Su, Q. (2010) Growth inhibitory effect and Chk1-dependent signaling involved in G2/M arrest on human gastric cancer cells induced by diallyl disulfide. Braz. J. Med. Biol. Res. 43, 271-278. https://doi.org/10.1590/S0100-879X2010007500004
  17. Cimprich, K. A. and Cortez, D. (2008) ATR: an essential regulator of genome integrity. Mol. Cell Biol. 9, 616-627.
  18. Parker, A., Gu, Y., Mahoney, W., Lee, S. H., Singh, K. K. and Lu, A. L. (2001) Human homolog of the MutY repair protein (hMYH) physically interacts with proteins involved in long patch DNA base excision repair. J. Biol. Chem. 276, 5547-5555. https://doi.org/10.1074/jbc.M008463200
  19. Gu, Y., Parker, A., Wilson, T. M., Bai, H., Chang, D. Y. and Lu, A. L. (2002) Human MutY homolog (hMYH), DNA glycosylase involved in base excision repair, physically and functionally interacts with mismatch repair proteins hMSH2/hMSH6. J. Biol. Chem. 277, 11135-11142. https://doi.org/10.1074/jbc.M108618200
  20. Yoshioka, K. I., Yoshioka, Y. and Hsieh, P. (2005) ATR kinase activation mediated by MutSα and MutLα in response to cytotoxic O6-methylguanine adducts. Molecular Cell 22, 501-510.
  21. Chang, D. Y. and Lu, A. L. (2005) Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. J. Biol. Chem. 280, 408-417. https://doi.org/10.1074/jbc.M406800200
  22. Lupardus, P. J. and Cimprich, K. A. (2006) Phosphorylation of Xenopus Rad1 and Hus1 defines a readout for ATR activation that is independent of claspin and the Rad9 carboxy terminus. Mol. Biol. Cell 17, 1559-1569. https://doi.org/10.1091/mbc.E05-09-0865
  23. Zou, L., Cortez, D. and Elledge, S. J. (2002) Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin. Genes Dev. 16, 198-208. https://doi.org/10.1101/gad.950302
  24. Bao, S., Lu, T., Wang, A., Zheng, H., Wang, L. E., Wei, Q., Hittelman, W. N. and Li, L. (2004) Disruption of the Rad9/Rad1/Hus1 (9-1-1) complex leads to checkpoint signaling and replication defects. Oncogene 23, 5586-5593. https://doi.org/10.1038/sj.onc.1207753

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