Requirement for C-terminal end of fibroblast growth factor receptor 4 in translocation of acidic fibroblast growth factor to cytosol and nucleus.
The ability of COS cells to bind and internalise acidic fibroblast growth factor (aFGF) was studied after transient transfection of the cells with wild-type and mutated fibroblast growth factor receptor 4. In one case the tyrosine kinase of the receptor was inactivated by a point mutation in the active site, whereas in other cases parts of the receptor were deleted to remove various parts of the cytoplasmic domain. In all cases the receptors were expressed at the cell surface at a high level and the cells bound labelled growth factor efficiently and internalised it by endocytosis. Translocation of externally added aFGF across cellular membranes to reach the cytosol and nucleus was measured as transport of labelled growth factor to the nuclear fraction obtained by centrifugation, by farnesylation of growth factor modified to carry a CAAX motif, and by phosphorylation of the growth factor at a site specific for protein kinase C. Whereas both full-length receptors (with and without an active kinase domain) facilitated translocation of the growth factor to the cytosol and nucleus, as assessed by these methods, the mutants of the receptor where the C terminus was deleted, were unable to do so. In contrast, a receptor containing only the 57 most C-terminal amino acids of the cytoplasmic domain in addition to the juxtamembrane, transmembrane and extracellular domains, was in fact able to mediate translocation of aFGF to the cytosol. These data indicate that information contained in the C terminus of the receptor is required for translocation.
Klingenberg
O
O
Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, Norway.
Wiedlocha
A
A
Rapak
A
A
Khnykin
D
D
Citores
L
L
Olsnes
S
S
D000818:Animals; D001665:Binding Sites / Q000502:physiology; D001692:Biological Transport / Q000502:physiology; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D004705:Endocytosis / Q000502:physiology; D016220:Fibroblast Growth Factor 1 / Q000378:metabolism*; D017353:Gene Deletion; D010770:Phosphotransferases / Q000378:metabolism; D017354:Point Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection
D002352:Carrier Proteins; D017468:Receptors, Fibroblast Growth Factor; D016220:Fibroblast Growth Factor 1; D010770:Phosphotransferases