Differential proteome analysis of replicative senescence in rat embryo fibroblasts

doi:10.1074/mcp.M100028-MCP200

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Submitted on October 31, 2001
Revised on March 11, 2002
Accepted on March 11, 2002

Differential proteome analysis of replicative senescence in rat embryo fibroblasts

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

Ludwig Institute for Cancer Research, London W1W 7BS

Corresponding Author: parmjit{at}ludwig.ucl.ac.uk

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 2-D differential proteome analysis of replicative senescence in serially passaged rat embryo fibroblasts was undertaken. Triplicate independent 2-D gels containing over 1200 spots each were run, curated and analysed. This revealed 49 spots whose expression was altered more than two-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 tumour suppressor for breast cancer and -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 ageing. Changes in expression of TUC-1, 2, 4 and 4 , 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 previously unrecognised in replicative senescence. They are now implicated in a new role.

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