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Papers In Press, published online ahead of print September 27, 2002
J. Biol. Chem, 10.1074/jbc.M207957200
Submitted on August 5, 2002
Revised on September 27, 2002
Accepted on September 27, 2002
in Rad18-independent post-replication repair of UV damage
Radiation Genetics, Kyoto University Graduate School of Medicine, Kyoto 606-8501
Corresponding Author: stakeda{at}rg.med.kyoto-u.ac.jp
DNA damages, which are left unrepaired by excision repair pathways, often block replication, leading to lesions such as breaks and gaps on the sister chromatids. These lesions may be processed by either homologous recombination (HR) repair or translesion DNA synthesis (TLS). Vertebrate Pol
belongs to the DNA polymerase Y family, as do most TLS polymerases. However, the role for Pol in vertebrate cells is unclear because of the lack of reverse genetic studies. Here, we generated cells deficient in Pol (pol cells) from the chicken B lymphocyte line DT40. Although purified Pol is unable to bypass ultraviolet (UV) damage, pol cells exhibited increased UV sensitivity and the phenotype was suppressed by expression of human and chicken Pol, suggesting that Pol is involved in TLS of UV photoproduct. Defects in both Pol and Rad18, which regulates TLS in yeast, in DT40 showed an additive effect on UV sensitivity. Interestingly, the level of sister chromatid exchange, which reflects HR-mediated repair, was elevated in normally cycling pol cells. This implies functional redundancy between HR and Pol in maintaining chromosomal DNA. In conclusion, vertebrate Pol is involved in Rad18-independent TLS of UV damage, and plays a role in maintaining genomic stability.
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