C3G is required for c-Abl-induced filopodia and its overexpression promotes filopodia formation.
The Rap1 guanine nucleotide exchange factor, C3G (also known as Rap1GEF-1) is involved in signaling from growth factors, cytokines and integrins and plays a role in cell adhesion and migration, but the mechanism by which C3G regulates various cellular functions is poorly understood. We, therefore, investigated the ability of C3G to affect actin cytoskeleton-dependent morphological changes in cells. Using RNA interference, we provide evidence that C3G is required for c-Abl-induced filopodia during cell spreading on fibronectin. C3G expression induces actin cytoskeletal reorganization and promotes filopodia formation independent of its catalytic activity. It showed enrichment at filopodia tips characteristic of molecules involved in filopodia dynamics. C3G-induced filopodia were not inhibited by dominant negative mutants of Rho, Rac and Cdc42, but required Abl catalytic activity. Coexpression of N-Wasp-Crib inhibited C3G induced as well as c-Abl-induced filopodia and wiskostatin, a pharmacological inhibitor of N-Wasp attenuates C3G-induced filopodia. Cellular C3G interacts with c-Abl and C3G expression results in enhanced localization of endogenous c-Abl in the cytoplasm. We suggest that C3G and c-Abl function in an interdependent manner, in linking external signals to remodeling the cytoskeleton to induce filopodia.
Radha
Vegesna
V
Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, India. vradha@ccmb.res.in
Rajanna
Ajumeera
A
Mitra
Aninda
A
Rangaraj
Nandini
N
Swarup
Ghanshyam
G
D000818:Animals; D020134:Catalytic Domain; D002448:Cell Adhesion / Q000235:genetics; D002460:Cell Line; D002465:Cell Movement / Q000235:genetics; D003593:Cytoplasm / Q000737:chemistry; D020735:Guanine Nucleotide-Releasing Factor 2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051304:Profilins / Q000378:metabolism; D016315:Proto-Oncogene Proteins c-abl / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011554:Pseudopodia / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000378:metabolism; D020764:cdc42 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism
D020735:Guanine Nucleotide-Releasing Factor 2; D051304:Profilins; D034741:RNA, Small Interfering; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D016315:Proto-Oncogene Proteins c-abl; D020764:cdc42 GTP-Binding Protein