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Nucleic Acids Research, 2000, Vol. 28, No. 18 3581-3586
© 2000 Oxford University Press

MDM2 interacts with the C-terminus of the catalytic subunit of DNA polymerase {varepsilon}

Nikolina Vlatkovic, Stephanie Guerrera, Ying Li1, Stuart Linn1, Dale S. Haines and Mark T. Boyd*

MCP Hahnemann University Cancer Center, Philadelphia, PA 19102, USA and 1Division of Biochemistry and Molecular Biology, University of California, Berkeley, CA 94720, USA

MDM2 is induced by p53 in response to cellular insults such as DNA damage and can have effects upon the cell cycle that are independent or downstream of p53. We used a yeast two-hybrid screen to identify proteins that bind to MDM2 and which therefore might be involved in these effects. We found that MDM2 can bind to the C-terminus of the catalytic subunit of DNA polymerase {varepsilon} (DNA pol {varepsilon}), to a region that is known to be essential in yeast. In an in vitro system we confirmed that MDM2 could bind to the homologous regions of both mouse and human DNA pol {varepsilon} and to full-length human DNA pol {varepsilon}. DNA pol {varepsilon} co-immunoprecipitated with MDM2 from transfected H1299 cells and also from a HeLa cell nuclear extract. We show here that the DNA pol {varepsilon}-interacting domain of MDM2 is located between amino acids 50 and 166. Our studies provide evidence that MDM2 interacts with a region of DNA pol {varepsilon} that plays a critical role in the function of DNA pol {varepsilon}.

* To whom correspondence should be addressed at present address: Department of Surgery, University of Liverpool, 5th Floor UCD, Daulby Street, Liverpool L69 3GA, UK. Tel: +44 151 706 4185; Fax: +44 151 706 5826; Email: mboyd@liverpool.ac.uk Present addresses: Nikolina Vlatkovic, University of Liverpool, Liverpool L69 3GA, UK Stephanie Guerrera, SmithKline-Beecham Pharmaceuticals, King of Prussia, PA 19406, USA Ying Li, Genetech, South San Francisco, CA 94080, USA Dale S. Haines, The Fels Institute for Cancer Research and Molecular Biology, Philadelphia, PA 19140, USA


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