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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

Ataxia telangiectasia and Rad3-related protein (ATR);Checkpoint kinase 1 (Chk1);Human MutY homolog (hMYH);Hydroxyurea (HU);Ultraviolet (UV)

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