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Mol Cell Biol, May 1998, p. 2758-2767, Vol. 18, No. 5
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Human CDC6/Cdc18 Associates with Orc1 and Cyclin-cdk and Is Selectively Eliminated from the Nucleus at the Onset of S Phase

Partha Saha, Junjie Chen,dagger Kelly C. Thome, Stephanie J. Lawlis, Zhi-hui Hou, Marvin Hendricks,Dagger Jeffrey D. Parvin, and Anindya Dutta*

Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115

Received 3 December 1997/Returned for modification 4 January 1998/Accepted 7 February 1998

In a two-hybrid screen for proteins that interact with human PCNA, we identified and cloned a human protein (hCdc18) homologous to yeast CDC6/Cdc18 and human Orc1. Unlike yeast, in which the rapid and total destruction of CDC6/Cdc18 protein in S phase is a central feature of DNA replication, the total level of the human protein is unchanged throughout the cell cycle. Epitope-tagged protein is nuclear in G1 and cytoplasmic in S-phase cells, suggesting that DNA replication may be regulated by either the translocation of this protein between the nucleus and the cytoplasm or the selective degradation of the protein in the nucleus. Mutation of the only nuclear localization signal of this protein does not alter its nuclear localization, implying that the protein is translocated to the nucleus through its association with other nuclear proteins. Rapid elimination of the nuclear pool of this protein after the onset of DNA replication and its association with human Orc1 protein and cyclin-cdks supports its identification as human CDC6/Cdc18 protein.

* Corresponding author. Mailing address: Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, 75 Francis St., Boston, MA 02115. Phone: (617) 278-0468. Fax: (617) 732-7449. E-mail: adutta{at}bustoff.bwh.harvard.edu.

dagger Present address: Cancer Biology Division, Dana-Farber Cancer Institute, Boston, MA 02115.

Dagger Present address: Millennium Pharmaceuticals Inc., Cambridge, MA 02139.

Mol Cell Biol, May 1998, p. 2758-2767, Vol. 18, No. 5
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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