Caveolin-1 polarization in transmigrating endothelial cells requires binding to intermediate filaments.
Caveolin-1 influences cell migration through multiple signaling pathways. In a previous report, we have shown that caveolin-1 is polarized in three-dimensional migrating endothelial cells (EC), and that caveolin-1 accumulation at the front of transmigrating cells requires the phosphorylatable Tyr(14) residue of caveolin-1. Immuno-electron microscopy further indicated that caveolin-1 was distributed along cytoskeletal structures in the anterior of transmigrating EC [Parat MO, Anand-Apte B, Fox PL (Mol Biol Cell 14:3156-3168, 2003)]. In the present study, we investigate whether caveolin-1 interacts with intermediate filaments (IF) and whether this interaction is required for caveolin-1 polarization in transmigrating cells. The distribution of vimentin is polarized in cells traversing a filter pore and overlaps with the distribution of caveolin-1, which accumulates in the cell front. In vivo sprouting EC also exhibit an anterior polarization of these two proteins. Furthermore, caveolin-1 co-purifies with intermediate filaments, suggesting an interaction between caveolin-1 and IF. Vimentin-deficient SW13 cells exhibit a dramatically altered polarization of caveolin-1-GFP, which no longer accumulates in the protruding cell extension. In addition, the Tyr(14) residue of caveolin-1 is required for co-purification of the protein with IF. Taken together, our results show that caveolin-1 Tyr(14) is necessary for binding to intermediate filaments, which in turn is required for anterior polarization of caveolin-1 in transmigrating cells.
Santilman
Virginie
V
Department of Anesthesiology Research, Cleveland Clinic, NE63, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
Baran
JoAnne
J
Anand-Apte
Bela
B
Evans
Robert M
RM
Parat
Marie-Odile
MO
D000818:Animals; D002417:Cattle; D051242:Caveolin 1 / Q000502:physiology*; D002465:Cell Movement; D016764:Cell Polarity; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D007382:Intermediate Filaments / Q000502:physiology; D014746:Vimentin / Q000502:physiology
D051242:Caveolin 1; D014746:Vimentin