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Vol. 11, Issue 6, 2057-2068, June 2000
*Department of Tumor Immunology, University Medical Center, NL-6525
EX Nijmegen, The Netherlands; and Restricted expression of activated leukocyte cell adhesion
molecule (ALCAM) by hematopoietic cells suggests an important role in
the immune system and hematopoiesis. To get insight into the mechanisms
that control ALCAM-mediated adhesion we have investigated homotypic
ALCAM-ALCAM interactions. Here, we demonstrate that the cytoskeleton
regulates ALCAM-mediated cell adhesion because inhibition of actin
polymerization by cytochalasin D (CytD) strongly induces homotypic
ALCAM-ALCAM interactions. This induction of cell adhesion is likely
due to clustering of ALCAM at the cell surface, which is observed after
CytD treatment. Single-particle tracking demonstrated that the lateral
mobility of ALCAM in the cell membrane is increased 30-fold after CytD
treatment. In contrast, both surface distribution and adhesion of a
glycosylphosphatidylinositol (GPI)-anchored ALCAM mutant are
insensitive to CytD, despite the increase in lateral mobility of
GPI-ALCAM upon CytD treatment. This demonstrates that clustering of
ALCAM is essential for cell adhesion, whereas enhanced diffusion of
ALCAM alone is not sufficient for cluster formation. In addition, upon
ligand binding, both free diffusion and the freely dragged distance of
wild-type ALCAM, but not of GPI-ALCAM, are reduced over time,
suggesting strengthening of the cytoskeleton linkage. From these
findings we conclude that activation of ALCAM-mediated adhesion is
dynamically regulated through actin cytoskeleton-dependent clustering.
Department of
Biophysical Techniques, University of Twente, NL-7500 AE
Enschede, The Netherlands
Online version of this article contains video
material for Figures 1-8. Online version available at
222.molbiolcell.org.
Corresponding author. E-mail address:
C.Figdor{at}mailbox.kun.nl.
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