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DDB1 gene disruption causes a severe growth defect and apoptosis in chicken DT40 cells |
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| Authors: | Wakasugi Mitsuo Matsuura Kenkyo Nagasawa Atsushi Fu DongTao Shimizu Hiroko Yamamoto Ken-ichi Takeda Shunichi Matsunaga Tsukasa |
| Institution: | aLaboratory of Human Molecular Genetics, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan;bDepartment of Molecular Pathology, Cancer Research Institute, Kanazawa University, 13-1 Takara-machi, Kanazawa 920-0934, Japan;cDepartment of Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan |
| Abstract: | DDB1 was originally identified as a heterodimeric complex with DDB2 and plays an accessory role in nucleotide excision repair. DDB1 also constitutes an E3 ubiquitin ligase complex together with Cul4A and Roc1 and acts as an adaptor, suggesting its multiple roles beyond DNA repair. We have generated a conditional DDB1-knockout mutant using a chicken B lymphocyte line DT40. Doxycycline-induced DDB1 depletion caused a severe growth defect followed by apoptotic cell death. Flow cytometric analyses revealed that cell cycle progression is initially retarded at all phases and subsequently impaired at S phase along with the appearance of sub-G1 population. Similarly, DDB1-knockdown in human U2OS cells by small interfering RNA exhibited a loss of clonogenic activity and perturbed cell cycle progression. These results demonstrate that the DDB1 gene is indispensable for cell viability in higher vertebrates and this conditional DDB1-knockout clone would be highly useful for the functional analysis of DDB1. |
| Keywords: | DDB1 DT40 Conditional knockout Cell viability Apoptosis Cell cycle arrest |
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