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PMID: 10648558 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

RhoA function in lamellae formation and migration is regulated by the alpha6beta4 integrin and cAMP metabolism.

The Journal of cell biology ·Vol. 148 ·No. 2 ·2000-01-24 ·Pages 253-8

O'Connor KL, Nguyen BK, Mercurio AM

Abstract

Clone A colon carcinoma cells develop fan-shaped lamellae and exhibit random migration when plated on laminin, processes that depend on the ligation of the alpha6beta4 integrin. Here, we report that expression of a dominant negative RhoA (N19RhoA) in clone A cells inhibited alpha6beta4-dependent membrane ruffling, lamellae formation, and migration. In contrast, expression of a dominant negative Rac (N17Rac1) had no effect on these processes. Using the Rhotekin binding assay to assess RhoA activation, we observed that engagement of alpha6beta4 by either antibody-mediated clustering or laminin attachment resulted in a two- to threefold increase in RhoA activation, compared with cells maintained in suspension or plated on collagen. Antibody-mediated clustering of beta1 integrins, however, actually suppressed Rho A activation. The alpha6beta4-mediated interaction of clone A cells with laminin promoted the translocation of RhoA from the cytosol to membrane ruffles at the edges of lamellae and promoted its colocalization with beta1 integrins, as assessed by immunofluorescence microscopy. In addition, RhoA translocation was blocked by inhibiting phosphodiesterase activity and enhanced by inhibiting the activity of cAMP-dependent protein kinase. Together, these results establish a specific integrin-mediated pathway of RhoA activation that is regulated by cAMP and that functions in lamellae formation and migration.

MeSH Terms
Antigens, Surface/isolation & purification,metabolism Cell Compartmentation Cell Membrane/metabolism,ultrastructure Cell Movement/physiology Collagen/metabolism Colonic Neoplasms/metabolism,ultrastructure Cyclic AMP/metabolism Cytoskeleton Humans Integrin alpha6beta4 Integrins/isolation & purification,metabolism Laminin/metabolism Tumor Cells, Cultured rhoA GTP-Binding Protein/isolation & purification,metabolism
Chemicals
Antigens, Surface Integrin alpha6beta4 Integrins Laminin laminin 1 Collagen Cyclic AMP rhoA GTP-Binding Protein
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
O'Connor K L
Division of Cancer Biology, Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
Nguyen B K
Mercurio A M
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.148.2.253
Published
2000-01-24
Pages
253-8
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2174290
Subset
IM
Grants
NCI NIH HHS · CA80789 · United States
External Links
PubMed source DOI: 10.1083/jcb.148.2.253 PMC Full Text View journal details
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