Mutants of basic fibroblast growth factor identify different cellular response programs.
Mutations, expected to affect the intracellular routing, i.e. additional nuclear localization sequences (NLS; the natural 23 kDa isoform and a 17D27R mutant) and/or a deletion of amino acids 26-29 (23 delta 26-29 and 17 delta 26-29), were introduced in basic fibroblast growth factor (bFGF). The mutants were assayed for their mitotic activity and their capacity to induce a tissue-specific response in human umbilical vein endothelial cells [HUVECs; induction of urokinase plasminogen activator receptor (u-PAR)], or in rat lens epithelial cells (fibre cell differentiation). In HUVECs, the 17D27R mutant had wild type activity, the 23 kDa and the delta 26-29 proteins were impaired in the induction of both mitosis and u-PAR. The delta 26-29 proteins, but not the 23 kDa protein or 17D27R mutant, were also impaired in receptor binding in that they bound only to a subset of receptors. The concentration of 17 kDa bFGF required for half maximal u-PAR response was 30 fold higher than for the half maximal 3H-thymidine incorporation. Addition of an NLS to bFGF strongly inhibited the induction of fibre cell differentiation, though it had little effect on the stimulation of DNA synthesis. The 17 delta 26-29 kDa mutant had wild type differentiation activity but was a poor mitogen for lens epithelial cells.
Leenders
W P
WP
Department of Molecular Biology and Cell Biology, University of Nijmegen, The Netherlands.
van Hinsbergh
V W
VW
van Genesen
S T
ST
Schoenmakers
J G
JG
van Zoelen
E J
EJ
Lubsen
N H
NH
D000818:Animals; D019556:COS Cells; D002454:Cell Differentiation; D002478:Cells, Cultured; D003459:Crystallins / Q000096:biosynthesis; D004247:DNA / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007908:Lens, Crystalline / Q000166:cytology / Q000378:metabolism; D008934:Mitogens / Q000502:physiology; D009154:Mutation / Q000502:physiology*; D019913:Nuclear Localization Signals / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D011956:Receptors, Cell Surface / Q000096:biosynthesis*; D055293:Receptors, Urokinase Plasminogen Activator; D011993:Recombinant Fusion Proteins; D017384:Sequence Deletion; D014471:Umbilical Veins
D003459:Crystallins; D008934:Mitogens; D019913:Nuclear Localization Signals; C524898:PLAUR protein, human; C524900:Plaur protein, rat; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011993:Recombinant Fusion Proteins; D016222:Fibroblast Growth Factor 2; D004247:DNA