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PMID: 12604587 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of a tight junction-associated guanine nucleotide exchange factor that activates Rho and regulates paracellular permeability.

The Journal of cell biology ·Vol. 160 ·No. 5 ·2003-03-03 ·Pages 729-40

Benais-Pont G, Punn A, Flores-Maldonado C, Eckert J, Raposo G, Fleming TP, Cereijido M, Balda MS, Matter K

Abstract

Rho family GTPases are important regulators of epithelial tight junctions (TJs); however, little is known about how the GTPases themselves are controlled during TJ assembly and function. We have identified and cloned a canine guanine nucleotide exchange factor (GEF) of the Dbl family of proto-oncogenes that activates Rho and associates with TJs. Based on sequence similarity searches and immunological and functional data, this protein is the canine homologue of human GEF-H1 and mouse Lfc, two previously identified Rho-specific exchange factors known to associate with microtubules in nonpolarized cells. In agreement with these observations, immunofluorescence of proliferating MDCK cells revealed that the endogenous canine GEF-H1/Lfc associates with mitotic spindles. Functional analysis based on overexpression and RNA interference in polarized MDCK cells revealed that this exchange factor for Rho regulates paracellular permeability of small hydrophilic tracers. Although overexpression resulted in increased size-selective paracellular permeability, such cell lines exhibited a normal overall morphology and formed fully assembled TJs as determined by measuring transepithelial resistance and by immunofluorescence and freeze-fracture analysis. These data indicate that GEF-H1/Lfc is a component of TJs and functions in the regulation of epithelial permeability.

MeSH Terms
Actin Cytoskeleton/metabolism,ultrastructure Animals Cell Communication/genetics Cell Membrane/metabolism,ultrastructure Cell Membrane Permeability/genetics Cells, Cultured Dogs Epithelial Cells/metabolism,ultrastructure Extracellular Matrix/genetics,metabolism Gene Expression Regulation/physiology Guanine Nucleotide Exchange Factors/genetics,isolation & purification Humans Immunohistochemistry Microscopy, Electron Microtubules/metabolism,ultrastructure Proto-Oncogene Proteins/genetics,isolation & purification RNA Interference/physiology Rho Guanine Nucleotide Exchange Factors Spindle Apparatus/metabolism,ultrastructure Tight Junctions/metabolism,ultrastructure rho GTP-Binding Proteins/metabolism
Chemicals
ARHGEF2 protein, human Arhgef2 protein, mouse Guanine Nucleotide Exchange Factors Proto-Oncogene Proteins Rho Guanine Nucleotide Exchange Factors rho GTP-Binding Proteins
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Benais-Pont Gaelle
Department of Cell Biology, University of Geneva, 1211 Geneva, Switzerland.
Punn Anu
Flores-Maldonado Catalina
Eckert Judith
Raposo Graca
Fleming Tom P
Cereijido Marcelino
Balda Maria S
Matter Karl
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200211047
Published
2003-03-03
Epub
2003-00-25
Pages
729-40
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2173357
Subset
IM
Grants
Wellcome Trust · United Kingdom
Medical Research Council · G0100558 · United Kingdom
Biotechnology and Biological Sciences Research Council · G18784 · United Kingdom
External Links
PubMed source DOI: 10.1083/jcb.200211047 PMC Full Text View journal details
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