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Originally published In Press as doi:10.1074/jbc.M102051200 on August 2, 2001

J. Biol. Chem., Vol. 276, Issue 38, 35644-35651, September 21, 2001
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Interactions in the Error-prone Postreplication Repair Proteins hREV1, hREV3, and hREV7*

Yoshiki MurakumoDagger §, Yukiko OguraDagger , Hideshi Ishii, Shin-ichiro Numata, Masatoshi IchiharaDagger , Carlo M. Croce, Richard Fishel, and Masahide TakahashiDagger

From the Dagger  Department of Pathology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan and the  Department of Microbiology and Immunology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107

Most mutations after DNA damage in yeast Saccharomyces cerevisiae are induced by error-prone translesion DNA synthesis employing scRev1 and DNA polymerase zeta  that consists of scRev3 and scRev7 proteins. Recently, the human REV1 (hREV1) and REV3 (hREV3) genes were identified, and their products were revealed to be involved in UV-induced mutagenesis, as observed for their yeast counterparts. Human REV7 (hREV7) was also cloned, and its product was found to interact with hREV3, but the biological function of hREV7 remained unknown. We report here the analyses of precise interactions in the human REV proteins. The interaction between hREV1 and hREV7 was identified by the yeast two-hybrid library screening using a bait of hREV7, which was confirmed by in vitro and in vivo binding assays. The homodimerization of hREV7 was also detected in the two-hybrid analysis. In addition, the precise domains for interaction between hREV7 and hREV1 or hREV3 and for hREV7 homodimerization were determined. Although hREV7 interacts with both hREV1 and hREV3, a stable complex formation of the three proteins was undetectable in vitro. These findings suggest the possibility that hREV7 might play an important role in regulating the enzymatic activities of hREV1 and hREV3 for mutagenesis in response to DNA damage.


* This work was supported by a grant-in-aid for COE (Center of Excellence) research from the Ministry of Education, Science, Sports and Culture of Japan.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 nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF357886.

§ To whom correspondence should be addressed. Tel.: 81-52-744-2093; Fax: 81-52-744-2098; E-mail: murakumo@med.nagoya-u.ac.jp.


Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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