c-Abl, Lamellipodin, and Ena/VASP proteins cooperate in dorsal ruffling of fibroblasts and axonal morphogenesis.
BACKGROUND
Tight regulation of cell motility is essential for many physiological processes, such as formation of a functional nervous system and wound healing. Drosophila Abl negatively regulates the actin cytoskeleton effector protein Ena during neuronal development in flies, and it has been postulated that this may occur through an unknown intermediary. Lamellipodin (Lpd) regulates cell motility and recruits Ena/VASP proteins (Ena, Mena, VASP, EVL) to the leading edge of cells. However, the regulation of this recruitment has remained unsolved.


RESULTS
Here we show that Lpd is a substrate of Abl kinases and binds to the Abl SH2 domain. Phosphorylation of Lpd positively regulates the interaction between Lpd and Ena/VASP proteins. Consistently, efficient recruitment of Mena and EVL to Lpd at the leading edge requires Abl kinases. Furthermore, transient Lpd phosphorylation by Abl kinases upon netrin-1 stimulation of primary cortical neurons positively correlates with an increase in Lpd-Mena coprecipitation. Lpd is also transiently phosphorylated by Abl kinases upon platelet-derived growth factor (PDGF) stimulation, regulates PDGF-induced dorsal ruffling of fibroblasts and axonal morphogenesis, and cooperates with c-Abl in an Ena/VASP-dependent manner.


CONCLUSIONS
Our findings suggest that Abl kinases positively regulate Lpd-Ena/VASP interaction, Ena/VASP recruitment to Lpd at the leading edge, and Lpd-Ena/VASP function in axonal morphogenesis and in PDGF-induced dorsal ruffling. Our data do not support the suggested negative regulatory role of Abl for Ena. Instead, we propose that Lpd is the hitherto unknown intermediary between Abl and Ena/VASP proteins.
Michael
Magdalene
M
Randall Division of Cell and Molecular Biophysics, King's College London, New Hunt's House, Guy's Campus, London SE1 1UL, UK.
Vehlow
Anne
A
Navarro
Christel
C
Krause
Matthias
M
D000818:Animals; D001369:Axons / Q000502:physiology*; D015153:Blotting, Western; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000502:physiology*; D004331:Drosophila melanogaster / Q000378:metabolism / Q000502:physiology; D005347:Fibroblasts / Q000502:physiology*; D047468:Immunoprecipitation; D009024:Morphogenesis / Q000502:physiology*; D009414:Nerve Growth Factors / Q000502:physiology; D000075388:Netrin-1; D010766:Phosphorylation; D010982:Platelet-Derived Growth Factor / Q000502:physiology; D016315:Proto-Oncogene Proteins c-abl / Q000502:physiology*; D025521:Tumor Suppressor Proteins / Q000502:physiology
D004268:DNA-Binding Proteins; C092744:ENA-VASP proteins; D009414:Nerve Growth Factors; D010982:Platelet-Derived Growth Factor; D025521:Tumor Suppressor Proteins; D000075388:Netrin-1; D016315:Proto-Oncogene Proteins c-abl