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PMID: 16055505 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mechanism of IFN-gamma-induced endocytosis of tight junction proteins: myosin II-dependent vacuolarization of the apical plasma membrane.

Molecular biology of the cell ·Vol. 16 ·No. 10 ·2005-10-00 ·Pages 5040-52

Utech M, Ivanov AI, Samarin SN, Bruewer M, Turner JR, Mrsny RJ, Parkos CA, Nusrat A

Abstract

Disruption of epithelial barrier by proinflammatory cytokines such as IFN-gamma represents a major pathophysiological consequence of intestinal inflammation. We have previously shown that IFN-gamma increases paracellular permeability in model T84 epithelial cells by inducing endocytosis of tight junction (TJ) proteins occludin, JAM-A, and claudin-1. The present study was designed to dissect mechanisms of IFN-gamma-induced endocytosis of epithelial TJ proteins. IFN-gamma treatment of T84 cells resulted in internalization of TJ proteins into large actin-coated vacuoles that originated from the apical plasma membrane and resembled the vacuolar apical compartment (VAC) previously observed in epithelial cells that lose cell polarity. The IFN-gamma dependent formation of VACs required ATPase activity of a myosin II motor but was not dependent on rapid turnover of F-actin. In addition, activated myosin II was observed to colocalize with VACs after IFN-gamma exposure. Pharmacological analyses revealed that formation of VACs and endocytosis of TJ proteins was mediated by Rho-associated kinase (ROCK) but not myosin light chain kinase (MLCK). Furthermore, IFN-gamma treatment resulted in activation of Rho GTPase and induced expressional up-regulation of ROCK. These results, for the first time, suggest that IFN-gamma induces endocytosis of epithelial TJ proteins via RhoA/ROCK-mediated, myosin II-dependent formation of VACs.

MeSH Terms
Actins/metabolism Adenosine Triphosphate/metabolism Cell Line Cell Membrane/physiology,ultrastructure Cell Polarity Claudin-1 Endocytosis Epithelial Cells/physiology,ultrastructure Interferon-gamma/pharmacology,physiology Intracellular Signaling Peptides and Proteins Membrane Proteins/metabolism Myosin Type II/physiology Myosin-Light-Chain Kinase/metabolism Occludin Protein Serine-Threonine Kinases/metabolism Recombinant Proteins Tight Junctions/physiology Up-Regulation Vacuoles/physiology rho GTP-Binding Proteins/metabolism rho-Associated Kinases
Chemicals
Actins Claudin-1 Intracellular Signaling Peptides and Proteins Membrane Proteins Occludin Recombinant Proteins Interferon-gamma Adenosine Triphosphate Protein Serine-Threonine Kinases rho-Associated Kinases Myosin-Light-Chain Kinase Myosin Type II rho GTP-Binding Proteins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Utech Markus
Epithelial Pathobiology Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322, USA.
Ivanov Andrei I
Samarin Stanislav N
Bruewer Matthias
Turner Jerrold R
Mrsny Randall J
Parkos Charles A
Nusrat Asma
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-03-0193
Published
2005-10-00
Epub
2005-00-29
Pages
5040-52
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1237102
Subset
IM
Grants
NIDDK NIH HHS · R01 DK059888 · United States
NIDDK NIH HHS · R01 DK061379 · United States
NIDDK NIH HHS · DK 59888 · United States
NIDDK NIH HHS · DK 61379 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-03-0193 PMC Full Text View journal details
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