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Published 25 October 2004. doi:10.1083/jcb.200403091
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 167, Number 2, 327-337
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Article

 Roles of p-ERM and Rho–ROCK signaling in lymphocyte polarity and uropod formation

Jong-Hwan Lee1, Tomoya Katakai1, Takahiro Hara1, Hiroyuki Gonda1,2, Manabu Sugai1, and Akira Shimizu1,2

1 Center for Molecular Biology and Genetics, Kyoto University
2 Translational Research Center, Kyoto University Hospital, Sakyo-ku, Kyoto 606-8507, Japan

Correspondence to Tomoya Katakai: tkatakai{at}virus.kyoto-u.ac.jp

Front–rear asymmetry in motile cells is crucial for efficient directional movement. The uropod in migrating lymphocytes is a posterior protrusion in which several proteins, including CD44 and ezrin/radixin/moesin (ERM), are concentrated. In EL4.G8 T-lymphoma cells, Thr567 phosphorylation in the COOH-terminal domain of ezrin regulates the selective localization of ezrin in the uropod. Overexpression of the phosphorylation-mimetic T567D ezrin enhances uropod size and cell migration. T567D ezrin also induces construction of the CD44-associated polar cap, which covers the posterior cytoplasm in staurosporine-treated, uropod-disrupted EL4.G8 cells or in naturally unpolarized X63.653 myeloma cells in an actin cytoskeleton–dependent manner. Rho-associated coiled coil–containing protein kinase (ROCK) inhibitor Y-27632 disrupts the uropod but not the polar cap, indicating that Rho–ROCK signaling is required for posterior protrusion but not for ERM phosphorylation. Phosphorylated ezrin associates with Dbl through its NH2-terminal domain and causes Rho activation. Moreover, constitutively active Q63L RhoA is selectively localized in the rear part of the cells. Thus, phosphorylated ERM has a potential function in establishing plasma membrane "posteriority" in the induction of the uropod in T lymphocytes.

J.-H. Lee and T. Katakai contributed equally to this work.

Abbreviations used in this paper: AID, autoinhibitory domain; BIM, bisindolylmaleimide I; CT, COOH-terminal; ERM, ezrin/radixin/moesin; GEF, guanine nucleotide exchange factor; MTOC, microtubule organizing center; NT, NH2-terminal; p-ERM, phosphorylated ERM; ROCK, Rho-associated coiled coil–containing protein kinase; SDF-1, stromal cell–derived factor-1; WT, wild-type.


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