Transformation of normal rat kidney cells by v-K-ras enhances expression of transin 2 and an S-100-related calcium-binding protein.
v-K-ras transformants of normal rat kidney cells (KNRK) exhibit cell surface-related, transformation-specific properties, including cell-surface fibronectin depletion, induction of anchorage- and density-independent growth, and increased synthesis of transforming growth factors alpha and beta. To search for potential distal effectors of v-K-ras-mediated transformation, we prepared a rabbit antiserum directed against intact KNRK cells to immunoprecipitate and compare proteins from detergent lysates and conditioned media of labeled NRK, KNRK, B77-NRK (a v-src transformant) and ts-371-NRK cells (a Ki-MSV encoding a temperature-sensitive p21v-K-ras). Proteins with enhanced expression in both wild-type v-K-ras and v-src transformants included a cell-surface phosphoglycoprotein with apparent Mr of 79,000 (79K) modified from an 85K protein observed in NRK cell lysates, a cytoplasmic 47K and a 10K protein, and a 57K secreted glycoprotein. A KNRK-specific 21K membrane-associated protein and secreted 59K and 36K secreted glycoproteins were also detected. The expression of the 36K and 59K proteins best correlated with temperature-dependent activation of the ts-371-NRK p21v-K-ras. Immunoselection of recombinant clones from a KNRK-specific lambda gt11 cDNA library allowed identification of the 59K and 10K proteins as transin 2 and an S-100-related calcium-binding protein identified as p9Ka/42A but not previously associated with oncogenic transformation of rat cells. Transin 2 detection by a cell-derived antiserum may also suggest the presence of specific cell-surface binding sites for this enzyme.
De Vouge
M W
MW
Department of Biology, McGill University, Montreal, Canada.
Mukherjee
B B
BB
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000235:genetics / Q000502:physiology*; D011905:Genes, ras; D006023:Glycoproteins / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry; D007668:Kidney / Q000166:cytology; D053506:Matrix Metalloproteinase 10; D008666:Metalloendopeptidases; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D051381:Rats; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins
D002135:Calcium-Binding Proteins; D006023:Glycoproteins; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; C499412:S100a4 protein, rat; D008666:Metalloendopeptidases; D053506:Matrix Metalloproteinase 10