Ligand-induced transphosphorylation between different FGF receptors.
Recent evidence shows that different fibroblast growth factors (FGF) bind with similar high affinities to two FGF receptors (FGFR) called flg and bek. In order to explore the mechanism of FGFR tyrosine autophosphorylation, we have generated cell lines which co-express a kinase-negative mutant of FGFR and an active form of FGFR. The following transfected NIH 3T3 cells were generated: (i) cells which express a shorter truncated form of bek (two Ig domains) together with a kinase-negative mutant of full length bek (bek K517A), (ii) cells which express wild-type bek together with kinase-negative flg (flg K514A) and (iii) cells co-expressing wild-type flg together with bek K517A. Immunoprecipitations with either bek-or flg-specific antisera followed by immunoblotting indicated that the double transfectants express the desired receptor species. The addition of acidic FGF (aFGF) to the various cell lines followed by immunoprecipitation with anti-FGFR antibodies and immunoblotting with anti-phosphotyrosine specific antibodies indicated that aFGF induces tyrosine phosphorylation of the kinase-negative FGFR mutants. These results show that tyrosine autophosphorylation of the kinase-negative FGFR is mediated by a transphosphorylation mechanism and that both homologous (bek----bek) and heterologous (bek----flg and flg----bek) transphosphorylation occurs in living cells. Recent evidence shows that tyrosine autophosphorylation of receptors with tyrosine kinase activities is essential for mediating interactions with signaling molecules. Therefore, heterologous transphosphorylation could amplify the response of cells to various forms of FGFs and their cognate receptors.
Bellot
F
F
Molecular Biology Department, Rhône-Poulenc Rorer Central Research, King of Prussia, PA 19406.
Crumley
G
G
Kaplow
J M
JM
Schlessinger
J
J
Jaye
M
M
Dionne
C A
CA
D001483:Base Sequence; D015153:Blotting, Western; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D000091344:Filaggrin Proteins; D005822:Genetic Vectors; D006801:Humans; D006921:Hygromycin B / Q000378:metabolism; D008024:Ligands; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011233:Precipitin Tests; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017468:Receptors, Fibroblast Growth Factor; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism
C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D008024:Ligands; D011956:Receptors, Cell Surface; D017468:Receptors, Fibroblast Growth Factor; D006921:Hygromycin B; D014443:Tyrosine; D005346:Fibroblast Growth Factors