Ornithine decarboxylase- and ras-induced cell transformations: reversal by protein tyrosine kinase inhibitors and role of pp130CAS.
We have found that overexpression of human ornithine decarboxylase (ODC) induces cell transformation in NIH 3T3 and Rat-1 cells (M. Auvinen, A. Paasinen, L. C. Andersson, and E. Hölttä, Nature (London) 360:355-358, 1992). The ODC-transformed cells display increased levels of tyrosine phosphorylation, in particular of a cluster of 130-kDa proteins. Here we show that one of the proteins with enhanced levels of tyrosine phosphorylation in ODC-overexpressing cells is the previously described p130 substrate of pp60v-src, known to associate also with v-Crk and designated p130CAS. We also studied the role of protein tyrosine phosphorylation in the ODC-induced cell transformation by exposing the cells to herbimycin A, a potent inhibitor of Src-family kinases, and to other inhibitors of protein tyrosine kinases. Treatment with the inhibitors reversed the phenotype of ODC-transformed cells to normal, with an organized, filamentous actin cytoskeleton. Coincidentally, the tyrosine hyperphosphorylation of p130 was markedly reduced, while the level of activity of ODC remained highly elevated. A similar reduction in pp130 phosphorylation and reversion of morphology by herbimycin A were observed in v-src- and c-Ha-ras-transformed cells. In addition, we show that expression of antisense mRNA for p130CAS resulted in reversion of the transformed phenotype of all these cell lines. An increased level of tyrosine kinase activity, not caused by c-Src or c-Abl, was further detected in the cytoplasmic fraction of ODC-transformed cells. Preliminary characteristics of this kinase are shown. These data indicate that p130CAS is involved in cell transformation by ODC, c-ras, and v-src oncogenes, raise the intriguing possibility that p130CAS may be generally required for transformation, and imply that there is at least one protein tyrosine kinase downstream of ODC that is instrumental for cell transformation.
Department of Pathology, Haartman Institute, University of Helsinki, Finland.
M
M
Auvinen
A
A
Paasinen-Sohns
H
H
Hirai
L C
LC
Andersson
E
E
Hölttä
D016475:3T3 Cells; D000199:Actins / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D016227:Benzoquinones; D015153:Blotting, Western; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D050719:Crk-Associated Substrate Protein; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D011905:Genes, ras; D006801:Humans; D047029:Lactams, Macrocyclic; D051379:Mice; D009955:Ornithine Decarboxylase / Q000096:biosynthesis* / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011506:Proteins; D011809:Quinones / Q000494:pharmacology*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism; D050717:Retinoblastoma-Like Protein p130; D017828:Rifabutin / Q000031:analogs & derivatives; D014162:Transfection; D019061:src-Family Kinases / Q000378:metabolism
D000199:Actins; C493841:BCAR1 protein, human; C493842:Bcar1 protein, mouse; C493843:Bcar1 protein, rat; D016227:Benzoquinones; D050719:Crk-Associated Substrate Protein; D004791:Enzyme Inhibitors; D047029:Lactams, Macrocyclic; D010750:Phosphoproteins; D011506:Proteins; D011809:Quinones; D011994:Recombinant Proteins; D016160:Retinoblastoma Protein; D050717:Retinoblastoma-Like Protein p130; D017828:Rifabutin; D019000:Phosphotyrosine; C020754:herbimycin; D011505:Protein-Tyrosine Kinases; D019061:src-Family Kinases; D009955:Ornithine Decarboxylase