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PMID: 16195341 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Zonula occludens-1 alters connexin43 gap junction size and organization by influencing channel accretion.

Molecular biology of the cell ·Vol. 16 ·No. 12 ·2005-12-00 ·Pages 5686-98

Hunter AW, Barker RJ, Zhu C, Gourdie RG

Abstract

Regulation of gap junction (GJ) organization is critical for proper function of excitable tissues such as heart and brain, yet mechanisms that govern the dynamic patterning of GJs remain poorly defined. Here, we show that zonula occludens (ZO)-1 localizes preferentially to the periphery of connexin43 (Cx43) GJ plaques. Blockade of the PDS95/dlg/ZO-1 (PDZ)-mediated interaction between ZO-1 and Cx43, by genetic tagging of Cx43 or by a membrane-permeable peptide inhibitor that contains the Cx43 PDZ-binding domain, led to a reduction of peripherally associated ZO-1 accompanied by a significant increase in plaque size. Biochemical data indicate that the size increase was due to unregulated accumulation of gap junctional channels from nonjunctional pools, rather than to increased protein expression or decreased turnover. Coexpression of native Cx43 fully rescued the aberrant tagged-connexin phenotype, but only if channels were composed predominately of untagged connexin. Confocal image analysis revealed that, subsequent to GJ nucleation, ZO-1 association with Cx43 GJs is independent of plaque size. We propose that ZO-1 controls the rate of Cx43 channel accretion at GJ peripheries, which, in conjunction with the rate of GJ turnover, regulates GJ size and distribution.

MeSH Terms
Animals Connexin 43/chemistry,genetics,metabolism Gap Junctions/physiology Gene Expression Regulation Glutathione Transferase/genetics,metabolism HeLa Cells Heart/physiology Humans Kinetics Membrane Proteins/genetics,physiology Phosphoproteins/genetics,physiology Rats Rats, Sprague-Dawley Recombinant Fusion Proteins/metabolism Zonula Occludens-1 Protein
Chemicals
Connexin 43 Membrane Proteins Phosphoproteins Recombinant Fusion Proteins TJP1 protein, human Tjp1 protein, rat Zonula Occludens-1 Protein Glutathione Transferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hunter Andrew W
Department of Cell Biology and Anatomy, Cardiovascular Developmental Biology Center, Medical University of South Carolina, Charleston, SC 29425, USA. [email protected]
Barker Ralph J
Zhu Ching
Gourdie Robert G
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-08-0737
Email
[email protected]
Published
2005-12-00
Epub
2005-00-29
Pages
5686-98
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1289413
Subset
IM
Grants
NHLBI NIH HHS · HL07260 · United States
NHLBI NIH HHS · HL36059 · United States
NICHD NIH HHS · HD39946 · United States
NHLBI NIH HHS · R01 HL056728 · United States
NHLBI NIH HHS · P01 HL036059 · United States
NHLBI NIH HHS · HL56728 · United States
NICHD NIH HHS · P01 HD039946 · United States
NHLBI NIH HHS · T32 HL007260 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-08-0737 PMC Full Text View journal details
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