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Mol Cell Biol, May 1998, p. 2748-2757, Vol. 18, No. 5
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Negative Regulation of DNA Replication by the Retinoblastoma Protein Is Mediated by Its Association with MCM7

Jacqueline M. Sterner,1,2 Susan Dew-Knight,1,2 Christine Musahl,3 Sally Kornbluth,1,4 and Jonathan M. Horowitz1,2,*

Departments of Molecular Cancer Biology,1 Microbiology,2 and Cell Biology,4 Duke University Medical Center, Durham, North Carolina 27710, and the Division of Biology, Universitat Konstanz, D 77434 Konstanz, Federal Republic of Germany3

Received 29 October 1997/Returned for modification 4 December 1997/Accepted 13 February 1998

A yeast two-hybrid screen was employed to identify human proteins that specifically bind the amino-terminal 400 amino acids of the retinoblastoma (Rb) protein. Two independent cDNAs resulting from this screen were found to encode the carboxy-terminal 137 amino acids of MCM7, a member of a family of proteins that comprise replication licensing factor. Full-length Rb and MCM7 form protein complexes in vitro, and the amino termini of two Rb-related proteins, p107 and p130, also bind MCM7. Protein complexes between Rb and MCM7 were also detected in anti-Rb immunoprecipitates prepared from human cells. The amino-termini of Rb and p130 strongly inhibited DNA replication in an MCM7-dependent fashion in a Xenopus in vitro DNA replication assay system. These data provide the first evidence that Rb and Rb-related proteins can directly regulate DNA replication and that components of licensing factor are targets of the products of tumor suppressor genes.

* Corresponding author. Present address: Department of Anatomy, Physiological Sciences, and Radiology, College of Veterinary Medicine North Carolina State University, Raleigh, NC 27606. Phone: (919) 515 4479. Fax: (919) 515 3044. E-mail: jon_horowitz{at}ncsu.edu.

Mol Cell Biol, May 1998, p. 2748-2757, Vol. 18, No. 5
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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