A novel fibroblast growth factor-1 (FGF1) mutant that acts as an FGF antagonist.
BACKGROUND
Crosstalk between integrins and FGF receptors has been implicated in FGF signaling, but the specifics of the crosstalk are unclear. We recently discovered that 1) FGF1 directly binds to integrin alphavbeta3, 2) the integrin-binding site and FGF receptor (FGFR) binding site are distinct, and 3) the integrin-binding-defective FGF1 mutant (R50E) is defective in inducing FGF signaling although R50E still binds to FGFR and heparin and induces transient ERK1/2 activation.


RESULTS
We tested if excess R50E affect DNA synthesis and cell survival induced by WT FGF1 in BaF3 mouse pro-B cells expressing human FGFR1. R50E suppressed DNA synthesis and cell proliferation induced by WT FGF1. We tested if WT FGF1 and R50E generate integrin-FGF1-FGFR ternary complex. WT FGF1 induced ternary complex formation (integrin-FGF-FGFR1) and recruitment of SHP-2 to the complex in NIH 3T3 cells and human umbilical endothelial cells, but R50E was defective in these functions. It has been reported that sustained ERK1/2 activation is integrin-dependent and crucial to cell cycle entry upon FGF stimulation. We thus determined the time-course of ERK1/2 activation induced by WT FGF1 and R50E. We found that WT FGF1 induced sustained activation of ERK1/2, but R50E was defective in this function.


CONCLUSIONS
Our results suggest that 1) R50E is a dominant-negative mutant, 2) Ternary complex formation is involved in FGF signaling, 3) The defect of R50E to bind to integrin may be directly related to the antagonistic action of R50E. Taken together, these results suggest that R50E has potential as a therapeutic in cancer.
Yamaji
Satoshi
S
Department of Dermatology, University of California Davis School of Medicine, Sacramento, California, United States of America.
Saegusa
Jun
J
Ieguchi
Katsuaki
K
Fujita
Masaaki
M
Mori
Seiji
S
Takada
Yoko K
YK
Takada
Yoshikazu
Y
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000166:cytology; D016220:Fibroblast Growth Factor 1 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D016023:Integrins / Q000378:metabolism; D051379:Mice; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D046912:Multiprotein Complexes; D050505:Mutant Proteins / Q000378:metabolism / Q000502:physiology; D041681:NIH 3T3 Cells; D011485:Protein Binding / Q000235:genetics; D015398:Signal Transduction
D016023:Integrins; D046912:Multiprotein Complexes; D050505:Mutant Proteins; D016220:Fibroblast Growth Factor 1; D004247:DNA; D048052:Mitogen-Activated Protein Kinase 3