Oncogenic activation of p59fyn tyrosine protein kinase by mutation of its carboxyl-terminal site of tyrosine phosphorylation, tyrosine 528.
As a result of alternative splicing, the Src-related tyrosine protein kinase p59fyn consists of two distinct isoforms termed FynB and FynT. Whereas the first product accumulates principally in brain, the second is expressed in hemopoietic cells, especially in T-lymphocytes. There is increasing evidence that the Fyn proteins are critical for normal functions of neuronal and lymphoid cells. To better understand the regulation of the catalytic function of p59fyn, we have tested the effects of mutating the major site of in vivo tyrosine phosphorylation, tyrosine 528, on the biological and biochemical properties of this enzyme. Our studies showed that a tyrosine 528-->phenylalanine (Y528F) mutation converted either Fyn isoform into a dominant oncoprotein, capable of full transformation of rodent fibroblasts. However, while both Y528F p59fynT and Y528F p59fynB were able to transform NIH 3T3 cells, activated FynT molecules were consistently more efficient at this process. It was also found that expression of wild-type p59fyn or kinase-defective Y528F Fyn molecules failed to provoke transformation of NIH 3T3 cells, implying that the transforming capabilities of Y528F Fyn relied on deregulated catalytic activity. Contrary to an earlier study (Cheng, S. H., Espino, P. C., Marshall, J., Harvey, R., Merrill, J., and Smith, A. E. (1991) J. Virol. 65, 170-179), these findings showed that mutation of the conserved carboxyl-terminal tyrosine residue markedly stimulated the catalytic function of p59fyn in vivo, implying that dephosphorylation of tyrosine 528 is sufficient to produce biologically relevant activation of the Fyn kinase. Moreover, our results provided further indication that the two Fyn isoforms possess distinct biochemical activities that may dictate functional differences in normal cell physiology.
Davidson
D
D
McGill Cancer Centre, Montréal, Quebec, Canada.
Fournel
M
M
Veillette
A
A
D016475:3T3 Cells; D000818:Animals; D002384:Catalysis; D002471:Cell Transformation, Neoplastic; D004789:Enzyme Activation; D051379:Mice; D009154:Mutation; D010649:Phenylalanine / Q000235:genetics; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D051076:Proto-Oncogene Proteins c-fyn; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*
D011518:Proto-Oncogene Proteins; D014443:Tyrosine; D010649:Phenylalanine; D011505:Protein-Tyrosine Kinases; C494960:Fyn protein, mouse; D051076:Proto-Oncogene Proteins c-fyn