Am J Physiol Cell Physiol AJP: Cell Physiology
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Am J Physiol Cell Physiol 292: C1775-C1786, 2007. First published January 17, 2007; doi:10.1152/ajpcell.00420.2006
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MUSCLE CELL BIOLOGY AND CELL MOTILITY

Obligatory role for phospholipase C-{gamma}1 in villin-induced epithelial cell migration

Yaohong Wang, Alok Tomar, Sudeep P. George, and Seema Khurana

Department of Physiology, University of Tennessee Health Science Center, Memphis, Tennessee

Submitted 4 August 2006 ; accepted in final form 2 January 2007

While there is circumstantial evidence to suggest a requirement for phospholipase C-{gamma}1 (PLC-{gamma}1) in actin reorganization and cell migration, few studies have examined the direct mechanisms that link regulators of the actin cytoskeleton with this crucial signaling molecule. This study was aimed to examine the role that villin, an epithelial cell-specific actin-binding protein, and its ligand PLC-{gamma}1 play in migration in intestinal and renal epithelial cell lines that endogenously or ectopically express human villin. Basal as well as epidermal growth factor (EGF)-stimulated cell migration was accompanied by tyrosine phosphorylation of villin and its association with PLC-{gamma}1. Inhibition of villin phosphorylation prevented villin-PLC-{gamma}1 complex formation as well as villin-induced cell migration. The absolute requirement for PLC-{gamma}1 in villin-induced cell migration was demonstrated by measuring cell motility in PLC-{gamma}1–/– cells and by downregulation of endogenous PLC-{gamma}1. EGF-stimulated direct interaction of villin with the Src homology domain 2 domain of PLC-{gamma}1 at the plasma membrane was demonstrated in living cells by using fluorescence resonance energy transfer. These results demonstrate that villin provides an important link between the activation of phosphoinositide signal transduction pathway and epithelial cell migration.

fluorescence resonance energy transfer; actin


Address for reprint requests and other correspondence: S. Khurana, Dept. of Physiology, The Univ. of Tennessee, Health Science Center, Nash 402, 894 Union Ave., Memphis, TN 38163 (e-mail: skhurana{at}utmem.edu)






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