Expression of senescence-induced protein WS3-10 in vivo and in vitro

In our efforts to characterize cellular senescence we have shown that the mRNA encoding WS3-10 protein is overexpressed in senescent human diploid fibroblasts (HDF) when compared with their younger counterparts, and that forced expression of the WS3-10 cDNA in young HDF results in suppression of calcium-dependent membrane currents, presumably due in part to the presence of a calcium binding domain within the WS3-10 protein. We have now expressed this protein in E. coli and have obtained affinity purified antibodies. Western blot analysis utilizing these antibodies showed that WS3-10 protein is also overexpressed in senescent HDF when compared to young HDF, and in normal fetal lung HDF when compared to SV40-transformed fetal lung HDF. HeLa cells do not express WS3-10 protein, In addition, we looked for WS3-10-related species in a variety of rat tissues. Analysis of WS3-10 immunologically related proteins in rat tissue extracts revealed two WS3-10 homologs, sized 22 kDa and 20 kDa. The latter presumably result from proteolytic removal of the C-terminal end of the 22 kDa polypeptide. The ratio between these polypeptides varies in a tissue-specific manner. Two proteins immunologically related to WS3-10 with sizes of 39 kDa and 91 kDa were present in rat spleen and skeletal muscle, respectively.