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.
Grigoriev
V G
VG
Department of Medicine, University of Arkansas for Medical Sciences, Little Rock 72205-5484, USA.
Thweatt
R
R
Moerman
E J
EJ
Goldstein
S
S
D000199:Actins / Q000378:metabolism; D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008297:Male; D008840:Microfilament Proteins; D008970:Molecular Weight; D009124:Muscle Proteins / Q000096:biosynthesis*; D009928:Organ Specificity; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis
D000199:Actins; D008840:Microfilament Proteins; D009124:Muscle Proteins; D011994:Recombinant Proteins; C058992:Tagln protein, mouse; C051615:transgelin