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Originally published as MBC in Press, 10.1091/mbc.E03-07-0468 on November 14, 2003

Vol. 15, Issue 2, 588-599, February 2004

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Gelsolin Mediates Collagen Phagocytosis through a Rac-dependent Step

Pamela D. Arora *, Michael Glogauer *, Andras Kapus {dagger}, David J. Kwiatkowski {ddagger}, and Christopher A. McCulloch * §

* Canadian Institutes of Health Research Group in Matrix Dynamics, University of Toronto, Toronto, Ontario, Canada M5S 3E2; {dagger} Department of Surgery, University Health Network, University of Toronto, Toronto, Ontario, Canada M5G 2C4; and {ddagger} Department of Hematology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115

Submitted July 3, 2003; Revised September 12, 2003; Accepted September 30, 2003
Monitoring Editor: Richard Assoian

The role of gelsolin, a calcium-dependent actin-severing protein, in mediating collagen phagocytosis, is not defined. We examined {alpha}2{beta}1 integrin-mediated phagocytosis in fibroblasts from wild-type (WT) and gelsolin knockout (Gsn-) mice. After initial contact with collagen beads, collagen binding and internalization were 60% lower in Gsn- than WT cells. This deficiency was restored by transfection with gelsolin or with {beta}1 integrin-activating antibodies. WT cells showed robust rac activation and increased [Ca2+]i during early contact with collagen beads, but Gsn- cells showed very limited responses. Transfected gelsolin in Gsn- cells restored rac activation after collagen binding. Transfection of Gsn- cells with active rac increased collagen binding to WT levels. Chelation of intracellular calcium inhibited collagen binding and rac activation, whereas calcium ionophore induced rac activation in WT and Gsn- cells. We conclude that the ability of gelsolin to remodel actin filaments is important for collagen-induced calcium entry; calcium in turn is required for rac activation, which subsequently enhances collagen binding to unoccupied {alpha}2{beta}1 integrins.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E03-07-0468. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E03-07-0468.

§ Corresponding author. E-mail address: christopher.mcculloch{at}utoronto.ca.




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