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PMID: 15800060 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.

Differential roles for actin polymerization and a myosin II motor in assembly of the epithelial apical junctional complex.

Molecular biology of the cell ·Vol. 16 ·No. 6 ·2005-06-00 ·Pages 2636-50

Ivanov AI, Hunt D, Utech M, Nusrat A, Parkos CA

Abstract

Differentiation and polarization of epithelial cells depends on the formation of the apical junctional complex (AJC), which is composed of the tight junction (TJ) and the adherens junction (AJ). In this study, we investigated mechanisms of actin reorganization that drive the establishment of AJC. Using a calcium switch model, we observed that formation of the AJC in T84 intestinal epithelial cells began with the assembly of adherens-like junctions followed by the formation of TJs. Early adherens-like junctions and TJs readily incorporated exogenous G-actin and were disassembled by latrunculin B, thus indicating dependence on continuous actin polymerization. Both adherens-like junctions and TJs were enriched in actin-related protein 3 and neuronal Wiskott-Aldrich syndrome protein (N-WASP), and their assembly was prevented by the N-WASP inhibitor wiskostatin. In contrast, the formation of TJs, but not adherens-like junctions, was accompanied by recruitment of myosin II and was blocked by inhibition of myosin II with blebbistatin. In addition, blebbistatin inhibited the ability of epithelial cells to establish a columnar phenotype with proper apico-basal polarity. These findings suggest that actin polymerization directly mediates recruitment and maintenance of AJ/TJ proteins at intercellular contacts, whereas myosin II regulates cell polarization and correct positioning of the AJC within the plasma membrane.

MeSH Terms
Actins/metabolism Adherens Junctions/chemistry,metabolism,ultrastructure Bridged Bicyclo Compounds, Heterocyclic/pharmacology Carbazoles/pharmacology Cell Line Cell Polarity/drug effects Epithelial Cells/cytology,metabolism,ultrastructure Heterocyclic Compounds, 4 or More Rings/pharmacology Humans Intestinal Mucosa/cytology Microvilli/drug effects,ultrastructure Models, Biological Molecular Motor Proteins/metabolism Myosin Type II/metabolism Polymers Propanolamines/pharmacology Thiazoles/pharmacology Thiazolidines Tight Junctions/chemistry,metabolism,ultrastructure
Chemicals
Actins Bridged Bicyclo Compounds, Heterocyclic Carbazoles Heterocyclic Compounds, 4 or More Rings Molecular Motor Proteins Polymers Propanolamines Thiazoles Thiazolidines wiskostatin blebbistatin Myosin Type II latrunculin B
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ivanov Andrei I
Epithelial Pathobiology Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322, USA. aiivano@emory.edu
Hunt Dirk
Utech Markus
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.e05-01-0043
Email
aiivano@emory.edu
Published
2005-06-00
Epub
2005-00-30
Pages
2636-50
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1142412
Subset
IM
Grants
NIDDK NIH HHS · R29 DK055679 · United States
NIDDK NIH HHS · DK-72564 · United States
NIDDK NIH HHS · R01 DK072564 · United States
NIDDK NIH HHS · DK-55679 · United States
NIDDK NIH HHS · R24 DK064399 · United States
NIDDK NIH HHS · DK-61379 · United States
NIDDK NIH HHS · DK-59888 · United States
NIDDK NIH HHS · R01 DK055679 · 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.e05-01-0043 PMC Full Text View journal details
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