GEF at work: Vav in protruding filopodia.
The Dbl family proto-oncogene vav is a nucleotide exchange factor for Rho family GTPases and is involved in triggering cytoskeletal changes contributing to the alterations of cell shape and motility, as well as in the induction of gene expression. In vitro and in vivo Vav is regulated by multiple tyrosine phosphorylation and binding to phosphatidylinositol phosphates. Although recruitment of Vav to the plasma membrane appears important for the activation of Vav function, there is little information on the precise subcellular localization of Vav in living cells. Employing live video fluorescence and immunoelectron microscopy, we show that GFP-tagged full-length Vav, and several mutants in which the N-terminal regulatory calponin homology (CH) domain has been deleted, specifically localize to the tips of filopodia. This localization was congruent with a high content of tyrosine phosphorylation in these regions. Consistent with earlier observations, mutants lacking the C-terminal SH domain region were unable to translocate to the filopodia tips. The enrichment in filopodial tips persisted despite their lateral movement but was dependent on forward growth. Upon retraction, the signal was rapidly lost, indicating that Vav undergoes a specific and transient translocation in response to actin-based, protrusive events in filopodia.
Kranewitter
W J
WJ
Institute of Molecular Biology, Department of Cell Biology, Austrian Academy of Sciences, Salzburg, Austria.
Danninger
C
C
Gimona
M
M
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002465:Cell Movement / Q000502:physiology; D048429:Cell Size / Q000502:physiology; D022081:Cell Surface Extensions / Q000502:physiology*; D003599:Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D015870:Gene Expression / Q000502:physiology; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D008545:Melanoma / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009154:Mutation / Q000502:physiology; D015513:Oncogene Proteins / Q000378:metabolism*; D010766:Phosphorylation; D051567:Proto-Oncogene Proteins c-vav; D011554:Pseudopodia / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000199:Actins; D020662:Guanine Nucleotide Exchange Factors; D015513:Oncogene Proteins; D051567:Proto-Oncogene Proteins c-vav; C496619:Vav1 protein, mouse