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PMID: 14528017 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Endocytosis of epithelial apical junctional proteins by a clathrin-mediated pathway into a unique storage compartment.

Molecular biology of the cell ·Vol. 15 ·No. 1 ·2004-01-00 ·Pages 176-88

Ivanov AI, Nusrat A, Parkos CA

Abstract

The adherens junction (AJ) and tight junction (TJ) are key regulators of epithelial polarity and barrier function. Loss of epithelial phenotype is accompanied by endocytosis of AJs and TJs via unknown mechanisms. Using a model of calcium depletion, we defined the pathway of internalization of AJ and TJ proteins (E-cadherin, p120 and beta-catenins, occludin, JAM-1, claudins 1 and 4, and ZO-1) in T84 epithelial cells. Proteinase protection assay and immunocytochemistry revealed orchestrated internalization of AJs and TJs into a subapical cytoplasmic compartment. Disruption of caveolae/lipid rafts did not prevent endocytosis, nor did caveolin-1 colocalize with internalized junctional proteins. Furthermore, AJ and TJ proteins did not colocalize with the macropinocytosis marker dextran. Inhibitors of clathrin-mediated endocytosis blocked internalization of AJs and TJs, and junctional proteins colocalized with clathrin and alpha-adaptin. AJ and TJ proteins were observed to enter early endosomes followed by movement to organelles that stained with syntaxin-4 but not with markers of late and recycling endosomes, lysosomes, or Golgi. These results indicate that endocytosis of junctional proteins is a clathrin-mediated process leading into a unique storage compartment. Such mechanisms may mediate the disruption of intercellular contacts during normal tissue remodeling and in pathology.

MeSH Terms
Adaptor Protein Complex alpha Subunits/metabolism Adherens Junctions/drug effects,metabolism,physiology Calcium/pharmacology Caveolin 1 Caveolins/metabolism,physiology Cell Polarity/physiology Cells, Cultured Clathrin/metabolism,physiology Cytoskeletal Proteins/metabolism,physiology Endocytosis/physiology Endosomes/metabolism,physiology Epithelial Cells/metabolism,physiology Golgi Apparatus/metabolism,physiology Humans Intestinal Mucosa/metabolism Intestines/physiology Membrane Microdomains/metabolism Membrane Proteins/isolation & purification,metabolism,physiology Microscopy, Fluorescence Qa-SNARE Proteins Tight Junctions/drug effects,metabolism,physiology Trans-Activators/metabolism,physiology beta Catenin
Chemicals
Adaptor Protein Complex alpha Subunits CAV1 protein, human CTNNB1 protein, human Caveolin 1 Caveolins Clathrin Cytoskeletal Proteins Membrane Proteins Qa-SNARE Proteins Trans-Activators beta Catenin Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ivanov Andrei I
Epithelial Pathobiology Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia 30322, USA. [email protected]
Nusrat Asma
Parkos Charles A
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e03-05-0319
Email
[email protected]
Published
2004-01-00
Epub
2003-00-03
Pages
176-88
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC307538
Subset
IM
Grants
NIDDK NIH HHS · R29 DK055679 · United States
NIDDK NIH HHS · DK 55679 · United States
NIDDK NIH HHS · DK 61379 · United States
NIDDK NIH HHS · R24 DK064399 · United States
NIDDK NIH HHS · R01 DK055679 · United States
NIDDK NIH HHS · DK 59888 · United States
NIDDK NIH HHS · DK 64399 · United States
NIDDK NIH HHS · R01 DK061379 · United States
NIDDK NIH HHS · R01 DK059888 · United States
External Links
PubMed source DOI: 10.1091/mbc.e03-05-0319 PMC Full Text View journal details
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