MET(PRC) mutations in the Ron receptor result in upregulation of tyrosine kinase activity and acquisition of oncogenic potential.
Ron and Met are structurally related receptor tyrosine kinases that elicit a complex biological response leading to invasive growth. Naturally occurring point mutations activate the Met kinase in papillary renal carcinomas (MET(PRC) mutations). By site-directed mutagenesis, we generated homologous amino acid substitutions in the Ron kinase domain and analyzed the biochemical and biological properties of the mutant receptors. Among the mutations studied, D(1232)H and M(1254)T displayed transforming activity in NIH3T3 cells, inducing focus formation and anchorage-independent growth. The D(1232)H and M(1254)T substitutions resulted in increased Ron autophosphorylation both in vivo and in vitro and constitutive binding to intracellular signal transducers. Both mutations yielded a dramatic increase in catalytic efficiency, indicating a direct correlation between kinase activity and oncogenic potential. Molecular modeling of the Ron D(1232)H mutation suggests that this single amino acid substitution favors the transition of the kinase from the inactive to the active state. These data demonstrate that point mutations can confer transforming activity to the Ron receptor and show that RON is a potential oncogene.
Williams
T A
TA
Department of Molecular Oncology, Institute for Cancer Research and Treatment (IRCC), University of Torino School of Medicine, Torino, Italy.
Longati
P
P
Pugliese
L
L
Gual
P
P
Bardelli
A
A
Michieli
P
P
D016475:3T3 Cells; D019943:Amino Acid Substitution; D000818:Animals; D002471:Cell Transformation, Neoplastic; D004789:Enzyme Activation; D051379:Mice; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009857:Oncogenes; D010766:Phosphorylation; D017354:Point Mutation; D011487:Protein Conformation; D019859:Proto-Oncogene Proteins c-met / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D015854:Up-Regulation
D011956:Receptors, Cell Surface; D014443:Tyrosine; D019859:Proto-Oncogene Proteins c-met; C080835:RON protein; D020794:Receptor Protein-Tyrosine Kinases