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PMID: 10601344 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Induction of cell scattering by expression of beta1 integrins in beta1-deficient epithelial cells requires activation of members of the rho family of GTPases and downregulation of cadherin and catenin function.

The Journal of cell biology ·Vol. 147 ·No. 6 ·1999-12-13 ·Pages 1325-40

Gimond C, van Der Flier A, van Delft S, Brakebusch C, Kuikman I, Collard JG, Fässler R, Sonnenberg A

Abstract

Adhesion receptors, which connect cells to each other and to the surrounding extracellular matrix (ECM), play a crucial role in the control of tissue structure and of morphogenesis. In this work, we have studied how intercellular adhesion molecules and beta1 integrins influence each other using two different beta1-null cell lines, epithelial GE11 and fibroblast-like GD25 cells. Expression of beta1A or the cytoplasmic splice variant beta1D, induced the disruption of intercellular adherens junctions and cell scattering in both GE11 and GD25 cells. In GE11 cells, the morphological change correlated with the redistribution of zonula occluden (ZO)-1 from tight junctions to adherens junctions at high cell confluency. In addition, the expression of beta1 integrins caused a dramatic reorganization of the actin cytoskeleton and of focal contacts. Interaction of beta1 integrins with their respective ligands was required for a complete morphological transition towards the spindle-shaped fibroblast-like phenotype. The expression of an interleukin-2 receptor (IL2R)-beta1A chimera and its incorporation into focal adhesions also induced the disruption of cadherin-based adhesions and the reorganization of ECM-cell contacts, but failed to promote cell migration on fibronectin, in contrast to full-length beta1A. This indicates that the disruption of cell-cell adhesion is not simply the consequence of the stimulated cell migration. Expression of beta1 integrins in GE11 cells resulted in a decrease in cadherin and alpha-catenin protein levels accompanied by their redistribution from the cytoskeleton-associated fraction to the detergent-soluble fraction. Regulation of alpha-catenin protein levels by beta1 integrins is likely to play a role in the morphological transition, since overexpression of alpha-catenin in GE11 cells before beta1 prevented the disruption of intercellular adhesions and cell scattering. In addition, using biochemical activity assays for Rho-like GTPases, we show that the expression of beta1A, beta1D, or IL2R-beta1A in GE11 or GD25 cells triggers activation of both RhoA and Rac1, but not of Cdc42. Moreover, dominant negative Rac1 (N17Rac1) inhibited the disruption of cell-cell adhesions when expressed before beta1. However, all three GTPases might be involved in the morphological transition, since expression of either N19RhoA, N17Rac1, or N17Cdc42 reversed cell scattering and partially restored cadherin-based adhesions in GE11-beta1A cells. Our results indicate that beta1 integrins regulate the polarity and motility of epithelial cells by the induction of intracellular molecular events involving a downregulation of alpha-catenin function and the activation of the Rho-like G proteins Rac1 and RhoA.

MeSH Terms
Actins/metabolism Alternative Splicing Animals Cadherins/metabolism Cell Adhesion Cell Line Cell Movement Cell Size Cytoskeletal Proteins/genetics,metabolism Cytoskeleton/metabolism Down-Regulation Enzyme Activation Epithelial Cells/cytology,enzymology,metabolism Gene Expression Integrin beta1/genetics,metabolism Ligands Mice Mutation/genetics Phenotype Receptors, Interleukin-2/genetics,metabolism Recombinant Fusion Proteins/chemistry,genetics,metabolism Solubility alpha Catenin rac1 GTP-Binding Protein/chemistry,genetics,metabolism rho GTP-Binding Proteins/chemistry,genetics,metabolism rhoA GTP-Binding Protein/chemistry,genetics,metabolism
Chemicals
Actins Cadherins Ctnna1 protein, mouse Cytoskeletal Proteins Integrin beta1 Ligands Receptors, Interleukin-2 Recombinant Fusion Proteins alpha Catenin rac1 GTP-Binding Protein rho GTP-Binding Proteins rhoA GTP-Binding Protein
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Gimond C
Division of Cell Biology, The Netherlands Cancer Institute, 1066 CX Amsterdam,The Netherlands.
van Der Flier A
van Delft S
Brakebusch C
Kuikman I
Collard J G
Fässler R
Sonnenberg A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.147.6.1325
Published
1999-12-13
Pages
1325-40
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2168093
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.147.6.1325 PMC Full Text View journal details
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