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J. Biol. Chem., Vol. 278, Issue 32, 30051-30056, August 8, 2003

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Cloning and Characterization of hCTF18, hCTF8, and hDCC1

HUMAN HOMOLOGS OF AN SACCHAROMYCES CEREVISIAE COMPLEX INVOLVED IN SISTER CHROMATID COHESION ESTABLISHMENT^*

Carolin J. Merkle , Larry M. Karnitz , John T. Henry-Sánchez ¶ and Junjie Chen  ||

From the Graduate Program in Tumor Biology, Mayo Graduate School, Rochester, Minnesota 55905, the Divisions of Oncology Research and Molecular Pharmacology and Therapeutics, Mayo Clinic, Rochester, Minnesota 55905, and the ^¶School of Medicine, University of Puerto Rico, San Juan, Puerto Rico 00936

A growing body of evidence suggests that establishment of sister chromatid cohesion is dependent on replication fork passage over a precohesion area. In Saccharomyces cerevisiae, this process involves an alternative replication factor C (RFC) complex that contains the four small RFC subunits as well as CTF18, CTF8, and DCC1. Here, we show that an evolutionarily conserved homologous complex exists in the nucleus of human cells. We demonstrate that hCTF18, hCTF8, and hDCC1 interact with each other as well as with the p38 subunit of RFC. This alternative RFC-containing complex interacts with proliferating cell nuclear antigen but not with the Rad9/Rad1/Hus1 complex, a proliferating cell nuclear antigen-like clamp involved in the DNA damage response. hCTF18 preferentially binds chromatin during S phase, suggesting a role during replication. Our data provide evidence for the existence of an alternative RFC complex with a probable role in mammalian sister chromatid cohesion establishment.

Received for publication, November 13, 2002 , and in revised form, May 21, 2003.

^* This work was supported in part by the National Institutes of Health T32 Institutional Training Grant 75926 in Tumor Biology and a predoctoral fellowship from the Mayo Foundation (to C. J. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Div. of Oncology Research, Guggenheim Bldg., Rm. 1306, Mayo Clinic, 200 First St. SW, Rochester, MN 55905. Tel.: 507-538-1545; Fax: 507-284-3906; E-mail: chen.junjie{at}mayo.edu.

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