Three determinants in ezrin are responsible for cell extension activity

M Martin, C Roy, P Montcourrier, A Sahuquet and P Mangeat

Centre National de la Recherche Scientifique, Universite Montpellier II, France.

The ERM proteins--ezrin, radixin, and moesin--are key players in membrane-cytoskeleton interactions. In insect cells infected with recombinant baculoviruses, amino acids 1-115 of ezrin were shown to inhibit an actin- and tubulin-dependent cell-extension activity located in ezrin C-terminal domain (ezrin310-586), whereas full-length ezrin1- 586 did not induce any morphological change. To refine the mapping of functional domains of ezrin, 30 additional constructs were overexpressed in Sf9 cells, and the resulting effect of each was qualitatively and semiquantitatively compared. The removal of amino acids 13-30 was sufficient to release a cell-extension phenotype. This effect was abrogated if the 21 distal-most C-terminal amino acids were subsequently deleted (ezrin31-565), confirming the existence of a head- to-tail regulation in the whole molecule. Surprisingly, the deletion in full-length ezrin of the same 21 amino acids provided strong cell- extension competence to ezrin1-565, and this property was recovered in N-terminal constructs as short as ezrin1-310. Within ezrin1-310, amino acid sequences 13-30 and 281-310 were important determinants and acted in cooperation to induce cytoskeleton mobilization. In addition, these same residues are part of a new actin-binding site characterized in vitro in ezrin N-terminal domain.

Volume 8, Issue 8, pp. 1543-1557, 08/01/1997
Copyright © 1997 by The American Society for Cell Biology

This article has been cited by other articles:

				B. Benayoun, S. Baghdiguian, A. Lajmanovich, M. Bartoli, N. Daniele, E. Gicquel, N. Bourg, F. Raynaud, M.-A. Pasquier, L. Suel, et al. NF-{kappa}B-dependent expression of the antiapoptotic factor c-FLIP is regulated by calpain 3, the protein involved in limb-girdle muscular dystrophy type 2A FASEB J, May 1, 2008; 22(5): 1521 - 1529. [Abstract] [Full Text] [PDF]

				F. A. Wald, A. S. Oriolo, A. Mashukova, N. L. Fregien, A. H. Langshaw, and P. J. I. Salas Atypical protein kinase C (iota) activates ezrin in the apical domain of intestinal epithelial cells J. Cell Sci., March 1, 2008; 121(5): 644 - 654. [Abstract] [Full Text] [PDF]

				D. Brockschnieder, H. Sabanay, D. Riethmacher, and E. Peles Ermin, A Myelinating Oligodendrocyte-Specific Protein That Regulates Cell Morphology J. Neurosci., January 18, 2006; 26(3): 757 - 762. [Abstract] [Full Text] [PDF]

				F. A. Wald, A. S. Oriolo, M. L. Casanova, and P. J.I. Salas Intermediate Filaments Interact with Dormant Ezrin in Intestinal Epithelial Cells Mol. Biol. Cell, September 1, 2005; 16(9): 4096 - 4107. [Abstract] [Full Text] [PDF]

				L. Zhu, Y. Liu, and J. G. Forte Ezrin oligomers are the membrane-bound dormant form in gastric parietal cells Am J Physiol Cell Physiol, June 1, 2005; 288(6): C1242 - C1254. [Abstract] [Full Text] [PDF]

				G. Bompard, M. Martin, C. Roy, F. Vignon, and G. Freiss Membrane targeting of protein tyrosine phosphatase PTPL1 through its FERM domain via binding to phosphatidylinositol 4,5-biphosphate J. Cell Sci., June 15, 2003; 116(12): 2519 - 2530. [Abstract] [Full Text] [PDF]

				T. C. Dickson, C. D. Mintz, D. L. Benson, and S. R.J. Salton Functional binding interaction identified between the axonal CAM L1 and members of the ERM family J. Cell Biol., June 24, 2002; 157(7): 1105 - 1112. [Abstract] [Full Text] [PDF]

				E Brault, A Gautreau, M Lamarine, I Callebaut, G Thomas, and L Goutebroze Normal membrane localization and actin association of the NF2 tumor suppressor protein are dependent on folding of its N-terminal domain J. Cell Sci., January 5, 2001; 114(10): 1901 - 1912. [Abstract] [PDF]

				C. Barret, C. Roy, P. Montcourrier, P. Mangeat, and V. Niggli Mutagenesis of the Phosphatidylinositol 4,5-Bisphosphate (PIP2) Binding Site in the NH2-Terminal Domain of Ezrin Correlates with Its Altered Cellular Distribution J. Cell Biol., November 20, 2000; 151(5): 1067 - 1080. [Abstract] [Full Text] [PDF]

				J. K. Locker, A. Kuehn, S. Schleich, G. Rutter, H. Hohenberg, R. Wepf, and G. Griffiths Entry of the Two Infectious Forms of Vaccinia Virus at the Plasma Membane Is Signaling-Dependent for the IMV but Not the EEV Mol. Biol. Cell, July 1, 2000; 11(7): 2497 - 2511. [Abstract] [Full Text]

				V. L. Bonilha, S. C. Finnemann, and E. Rodriguez-Boulan Ezrin Promotes Morphogenesis of Apical Microvilli and Basal Infoldings in Retinal Pigment Epithelium J. Cell Biol., December 27, 1999; 147(7): 1533 - 1548. [Abstract] [Full Text] [PDF]

				Y. Doi, M. Itoh, S. Yonemura, S. Ishihara, H. Takano, T. Noda, S. Tsukita, and S. Tsukita Normal Development of Mice and Unimpaired Cell Adhesion/Cell Motility/Actin-based Cytoskeleton without Compensatory Up-regulation of Ezrin or Radixin in Moesin Gene Knockout J. Biol. Chem., January 22, 1999; 274(4): 2315 - 2321. [Abstract] [Full Text] [PDF]

				A Skoudy, G. Nhieu, N Mantis, M Arpin, J Mounier, P Gounon, and P Sansonetti A functional role for ezrin during Shigella flexneri entry into epithelial cells J. Cell Sci., January 7, 1999; 112(13): 2059 - 2068. [Abstract] [PDF]

				M Gronholm, M Sainio, F Zhao, L Heiska, A Vaheri, and O Carpen Homotypic and heterotypic interaction of the neurofibromatosis 2 tumor suppressor protein merlin and the ERM protein ezrin J. Cell Sci., January 3, 1999; 112(6): 895 - 904. [Abstract] [PDF]

				M. Amieva, P Litman, L Huang, E Ichimaru, and H Furthmayr Disruption of dynamic cell surface architecture of NIH3T3 fibroblasts by the N-terminal domains of moesin and ezrin: in vivo imaging with GFP fusion proteins J. Cell Sci., January 1, 1999; 112(1): 111 - 125. [Abstract] [PDF]

				G. Paglini, P. Kunda, S. Quiroga, K. Kosik, and A. Caceres Suppression of Radixin and Moesin Alters Growth Cone Morphology, Motility, and Process Formation In Primary Cultured Neurons J. Cell Biol., October 19, 1998; 143(2): 443 - 455. [Abstract] [Full Text] [PDF]

				T. Matsui, M. Maeda, Y. Doi, S. Yonemura, M. Amano, K. Kaibuchi, S. Tsukita, and S. Tsukita Rho-Kinase Phosphorylates COOH-terminal Threonines of Ezrin/Radixin/Moesin (ERM) Proteins and Regulates Their Head-to-Tail Association J. Cell Biol., February 9, 1998; 140(3): 647 - 657. [Abstract] [Full Text] [PDF]

				A. Murthy, C. Gonzalez-Agosti, E. Cordero, D. Pinney, C. Candia, F. Solomon, J. Gusella, and V. Ramesh NHE-RF, a Regulatory Cofactor for Na+-H+ Exchange, Is a Common Interactor for Merlin and ERM (MERM) Proteins J. Biol. Chem., January 16, 1998; 273(3): 1273 - 1276. [Abstract] [Full Text] [PDF]

				A Bretscher, D Reczek, and M Berryman Ezrin: a protein requiring conformational activation to link microfilaments to the plasma membrane in the assembly of cell surface structures J. Cell Sci., January 12, 1997; 110(24): 3011 - 3018. [Abstract] [PDF]