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J. Biol. Chem., Vol. 277, Issue 12, 10400-10409, March 22, 2002
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From the Intercellular adhesion molecule 3 (ICAM-3) is a
leukocyte-specific receptor involved in primary immune
responses. We have investigated the interaction between ICAM-3 and
ezrin/radixin/moesin (ERM) proteins and its role in LFA-1-induced
cell-cell interactions and membrane positioning of ICAM-3 in polarized
migrating lymphocytes. Protein-protein binding assays demonstrated a
phosphatidylinositol 4,5-bisphosphate-induced association between
ICAM-3 and the amino-terminal domain of ERM proteins. This interaction
was not essential for the binding of ICAM-3 to LFA-1. Dynamic
fluorescence videomicroscopy studies of cells demonstrated that moesin
and ICAM-3 coordinately redistribute on the plasma membrane during
lymphocyte migration. Furthermore, overexpression of the amino-terminal
domain of moesin, which lacks the consensus moesin actin-binding site,
caused the subcellular mislocalization of ICAM-3. A CD4 chimerical
protein containing the cytoplasmic tail of ICAM-3 was targeted to the trailing edge. Point mutation of Ser487,
Ser489, and Ser496 to alanine in the
juxtamembrane region of ICAM-3 significantly impaired both ERM binding
and polarization of ICAM-3. ERM-directed polarization of ICAM-3
was also impaired by phosphorylation-like mutation of
Ser487 and Ser489, but not of
Ser496. Our results underscore the key role of specific
serine residues within the cytoplasmic region of ICAM-3 for its
ERM-directed positioning at the trailing edge of motile lymphocytes.
A Novel Serine-rich Motif in the Intercellular Adhesion
Molecule 3 Is Critical for Its Ezrin/Radixin/Moesin-directed
Subcellular Targeting*
,,,,
,**
Servicio de Inmunología,
Hospital de la Princesa, Universidad Autónoma de Madrid, Madrid
28006, Spain, the Department of Pathology, Stanford University,
Stanford, California 94304, the ¶ Tumor Immunology Program, German
Cancer Research Center, D-69120 Heidelberg, Germany, and
§ Servei d'Immunologia, Hospital Clínic,
Barcelona 08036, Spain
*
This work was supported by Grants SAF99-0034-C01 and FEDER
2FD97-068-C02-02 from Ministerio de Educación, Grant
08.3/0010.1/99 from Comunidad Autónoma de Madrid, Grant
QLRT-1999-01036 from the European Community, and a grant from the
Tobacco-Related Disease Research Program of the State of
California (to H. F.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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