Effects of mutations of a phosphorylation site in an exposed loop in acidic fibroblast growth factor.
Acidic fibroblast growth factor (aFGF) contains a phosphorylation site recognized by protein kinase C. A non-mitogenic mutant growth factor is devoid of this phosphorylation site. We have changed amino acids in and close to the phosphorylation site and studied the consequences of this for binding of the growth factor to high affinity receptors as well as to heparin. We have also studied the ability of the mutants to stimulate DNA synthesis and cell proliferation as well as phosphorylation of mitogen-activated protein kinase and the ability of the growth factor mutants to be transported to the nucleus. The results indicate that while the mutations strongly affect the ability of the growth factor to bind to heparin, they do not affect much the binding to the specific FGF receptors, activation of mitogen-activated protein kinase or transport of the growth factor to the nucleus. The mutations affect to various extents the ability of the growth factor to stimulate DNA synthesis and to induce cell multiplication. We find that phosphorylation of aFGF is not required for mitogenic activity. The data suggest that altered interaction of the growth factor with a cellular component different from the receptor, possibly a component in the nucleus, is the reason for the different mitogenicity of the different growth factor mutants.
Klingenberg
O
O
Department of Biochemistry at The Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, 0310 Oslo, Norway.
Wiedlocha
A
A
Olsnes
S
S
D016475:3T3 Cells; D000818:Animals; D001665:Binding Sites; D001692:Biological Transport; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism; D004261:DNA Replication / Q000235:genetics; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry / Q000235:genetics*; D006493:Heparin / Q000378:metabolism; D051379:Mice; D009154:Mutation; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism
D017468:Receptors, Fibroblast Growth Factor; D016220:Fibroblast Growth Factor 1; D006493:Heparin; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases