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Originally published In Press as doi:10.1074/jbc.M207957200 on September 27, 2002

J. Biol. Chem., Vol. 277, Issue 50, 48690-48695, December 13, 2002
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Involvement of Vertebrate Polkappa in Rad18-independent Postreplication Repair of UV Damage*,

Takashi OkadaDagger §, Eiichiro SonodaDagger , Yukiko M. YamashitaDagger , Shogo KoyoshiDagger , Satoshi Tateishi||, Masaru Yamaizumi||, Minoru Takata**, Osamu Ogawa§, and Shunichi TakedaDagger Dagger Dagger

From the Dagger  Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Kyoto 606-8501, Japan, the § Department of Urology, Kyoto University Graduate School of Medicine, Kyoto 606-8507, Japan, the || Institute of Molecular Embryology and Genetics, Kumamoto University School of Medicine, Kumamoto 862-0976, Japan, and the ** Department of Immunology and Molecular Genetics, Kawasaki Medical College School of Medicine, Kurashiki 701-0192, Japan

DNA damage, which is left unrepaired by excision repair pathways, often blocks 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 Polkappa belongs to the DNA polymerase Y family, as do most TLS polymerases. However, the role for Polkappa in vertebrate cells is unclear because of the lack of reverse genetic studies. Here, we generated cells deficient in Polkappa (polkappa cells) from the chicken B lymphocyte line DT40. Although purified Polkappa is unable to bypass ultraviolet (UV) damage, polkappa cells exhibited increased UV sensitivity, and the phenotype was suppressed by expression of human and chicken Polkappa , suggesting that Polkappa is involved in TLS of UV photoproduct. Defects in both Polkappa 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 polkappa cells. This implies functional redundancy between HR and Polkappa in maintaining chromosomal DNA. In conclusion, vertebrate Polkappa is involved in Rad18-independent TLS of UV damage and plays a role in maintaining genomic stability.

* This work was supported by a grant from the Core Research for Evolution Science and Technology (CREST) of Japan Science and Technology Corporation, by a Center of Excellence (COE) grant from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by grants from the Uehara Memorial Foundation and the Mochida Memorial Foundation for Medical and Pharmaceutical Research.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental data.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AY118271.

Present address: Dept. of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305-5329.

Dagger Dagger To whom correspondence should be addressed: Dept. of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501. Japan. Tel.: 81-75-753-4410; Fax: 81-75-753-4419; E-mail: stakeda@rg.med.kyoto-u.ac.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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