Mutants of Basic Fibroblast Growth Factor Identify Different Cellular Response Programs

Abstract

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 (23A26–29 and 17Δ26–29), 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 Δ26–29 proteins were impaired in the induction of both mitosis and u-PAR. The Δ26–29 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 17Δ26–29 kDa mutant had wild type differentiation activity but was a poor mitogen for lens epithelial cells.

Key Words:

• bFGF mutants
• lens
• differentiation
• nuclear localization