Activated SRC oncogene phosphorylates R-ras and suppresses integrin activity.
One of the prominent effects of the Src kinase is to reduce cell adhesion. The small GTPase, R-Ras, affects cell adhesion by maintaining integrin activity, and the ability of R-Ras to do so can be regulated by phosphorylation of a tyrosine residue located in its effector domain by an Eph receptor kinase (Zou, J. X., Wang, B., Kalo, M. S., Zisch, A. H., Pasquale, E. B., and Ruoslahti, E. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 13813-13818). Here we show that Src regulates cell adhesion through R-Ras and integrins. Reduced substrate attachment of 293T cells transfected with the cDNA for an activated form of Src (v-Src) was accompanied by phosphorylation of endogenous R-Ras. v-Src also phosphorylated R-Ras in vitro. An activated form of Src similar to one that has been found in human cancers, Src527, shared with v-Src the ability to phosphorylate R-Ras. Stronger R-Ras phosphorylation was seen in Madin-Darby canine kidney cells cells transformed with temperature-sensitive v-Src at the permissive temperature than at the non-permissive temperature, and R-Ras and Src co-immunoprecipitated at the permissive temperature. Mutation analysis showed that the Src phosphorylation site in R-Ras was tyrosine 66, the position critical to the ability of R-Ras to support integrin activity. Finally, activated R-Ras in which tyrosine 66 is mutated to phenylalanine rendered cells partially resistant to the effects of Src on cell adhesion. Regulation of cell adhesion by Src through R-Ras may be at least partially responsible for the reduced adhesion and the resulting increased invasiveness of Src-transformed cells.
Zou
June X
JX
Cancer Research Center, The Burnham Institute, La Jolla, California 92037, USA.
Liu
Yanqiu
Y
Pasquale
Elena B
EB
Ruoslahti
Erkki
E
D000818:Animals; D002461:Cell Line, Transformed; D004285:Dogs; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D016391:Genes, src; D016023:Integrins / Q000037:antagonists & inhibitors*; D009361:Neoplasm Invasiveness; D010766:Phosphorylation; D018631:ras Proteins / Q000378:metabolism*
D016023:Integrins; D020558:GTP Phosphohydrolases; D018631:ras Proteins