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Vanadate‐treated baby hamster kidney fibroblasts show cytoskeleton and adhesion patterns similar to their rous sarcoma virus‐transformed counterparts
Author(s) -
Marchisio Pier Carlo,
D'Urso Nicoletta,
Comoglio Paolo M.,
Giancotti Filippo G.,
Tarone Guido
Publication year - 1988
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240370203
Subject(s) - podosome , rous sarcoma virus , baby hamster kidney cell , microbiology and biotechnology , cytoskeleton , tyrosine phosphorylation , vanadate , tyrosine , biology , phosphorylation , tyrosine kinase , cell adhesion , actin cytoskeleton , protein tyrosine phosphatase , actin , proto oncogene tyrosine protein kinase src , biochemistry , cell , signal transduction , immunology , virus
Rous sarcoma virus‐trans formed baby hamster kidney fibroblasts (RSV/B4‐BHK) adhere to a fibronectin‐coated substratum by means of dot‐like adhesion sites called podosomes in view of their shape and function as cellular feet (Tarone et al.: Exp Cell Res 159:141, 1985). Podosomes concentrate tyrosine‐phosphorylated proteins, including pp60 v‐src , and appear in many cells transformed by oncogenes coding for tyrosine kinases. In this paper we used orthovanadate, an inhibitor of phosphotyrosine phosphatases, in order to increase the cellular concentration of phosphotyrosine and to study whether this treatment induced the cytoskeleton remodeling leading to the formation of podosomes. Indeed, orthovanadate (10–100 μM) induced in a time‐and dose‐dependent manner the redistribution of F‐actin and the formation of podosomes in BHK cells. Cytoskeleton remodeling occurred along with a marked increase of tyrosine phosphorylatcd proteins. The vanadate effect on the cytoskeletal phenotype was enhanced by the simultaneous treatment of cells with a phorbol ester. Under the latter conditions almost all BHK cells showed podosomes. The vanadate effect was reversible insofar as podosomes and tyrosine‐phosphorylated proteins disappeared. Then, vanadate treatment of normal cells induced the cascade of events leading to the cytoskeletal changes typical of transformation and suggested that the transformed cytoskeletal phenotype may he primarily induced by the tyrosine phosphorylation of unknown target(s) operated by endogenous kinases.