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Originally published In Press as doi:10.1074/mcp.M100028-MCP200 on March 11, 2002.
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Molecular & Cellular Proteomics 1:280-292, 2002.
© 2002 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Differential Proteome Analysis of Replicative Senescence in Rat Embryo Fibroblasts*,S

Silvia Benvenuti{ddagger},§, Rainer Cramer{ddagger},§, Christopher C. Quinn, Jim Bruce||, Marketa Zvelebil{ddagger},§, Steven Corless{ddagger}, Jacquelyn Bond{ddagger}, Alice Yang{ddagger}, Susan Hockfield, Alma L. Burlingame{ddagger},§, Michael D. Waterfield{ddagger},§ and Parmjit S. Jat{ddagger},§,**

{ddagger} Ludwig Institute for Cancer Research, Royal Free and University College School of Medicine, Courtauld Building, 91 Riding House Street, London W1W 7BS, United Kingdom
§ Department of Biochemistry and Molecular Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom
Section of Neurobiology, Yale University School of Medicine, New Haven, Connecticut 06520-8001
|| Oxford GlycoSciences, 10 The Quadrant, Abingdon Science Park, Oxfordshire OX14 3YS, United Kingdom

Normal somatic cells undergo a finite number of divisions and then cease dividing whereas cancer cells are able to proliferate indefinitely. To identify the underlying mechanisms that limit the mitotic potential, a two-dimensional differential proteome analysis of replicative senescence in serially passaged rat embryo fibroblasts was undertaken. Triplicate independent two-dimensional gels containing over 1200 spots each were run, curated, and analyzed. This revealed 49 spots whose expression was altered more than 2-fold. Of these, 42 spots yielded positive protein identification by mass spectrometry comprising a variety of cytoskeletal, heat shock, and metabolic proteins, as well as proteins involved in trafficking, differentiation, and protein synthesis, turnover, and modification. These included gelsolin, a candidate tumor suppressor for breast cancer, and {alpha}-glucosidase II, a member of the family of glucosidases that includes klotho; a defect in klotho expression in mice results in a syndrome that resembles human aging. Changes in expression of TUC-1, -2, -4, and -4ß, members of the TUC family critical for neuronal differentiation, were also identified. Some of the identified changes were also shown to occur in two other models of senescence, premature senescence of REF52 cells and replicative senescence of mouse embryo fibroblasts. The majority of these candidate proteins were unrecognized previously in replicative senescence. They are now implicated in a new role.

** To whom correspondence should be addressed. Tel.: 44-2078784101; Fax: 44-2078784040; E-mail: parmjit{at}ludwig.ucl.ac.uk


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