A dominant negative mutation of neuronal connexin 36 that blocks intercellular permeability. | |
MedLine Citation:
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PMID: 11834292 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Rat connexin 36 (Cx36) was mutated by substituting serine for cysteine at residue 231 (C231S) and the mutant's effect on the subcellular localization of wild-type Cx36 and the intercellular permeability that it confers was determined in human HeLa and rat PC12 cells. Cells transfected with the mutant or wild-type Cx36 cDNA expressed the expected 36 kDa protein and Cx36 immunoreactivity. Co-immunoprecipitation experiments with monkey COS-7 cells transiently transfected with both mutant and wild-type Cx36 cDNAs demonstrated that the mutant protein bound to the wild-type. Double immunofluorescence microscopy of stably transfected HeLa cells demonstrated that mutant Cx36 blocked the transport of the wild-type Cx36 to the cell membrane, primarily by trapping it in the endoplasmic reticulum around the nucleus. Coexpression of the mutant Cx36 with the wild-type protein abolished the ability of the latter to permit dye transfer in both HeLa and PC12 cells. The findings are the first demonstration of a mutation of Cx36 that inhibits wild-type Cx36 function in mammalian cells. |
Authors:
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Dimitris Placantonakis; Federico Cicirata; John P Welsh |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Brain research. Molecular brain research Volume: 98 ISSN: 0169-328X ISO Abbreviation: Brain Res. Mol. Brain Res. Publication Date: 2002 Jan |
Date Detail:
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Created Date: 2002-02-08 Completed Date: 2002-04-29 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 8908640 Medline TA: Brain Res Mol Brain Res Country: Netherlands |
Other Details:
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Languages: eng Pagination: 15-28 Citation Subset: IM |
Affiliation:
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Department of Physiology and Neuroscience, New York University School of Medicine, 550 First Avenue, New York, NY 10016, USA. |
Export Citation:
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MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Substitution* Animals COS Cells / metabolism Cell Communication / genetics* Cercopithecus aethiops Connexins / genetics*, physiology DNA, Complementary / genetics Endoplasmic Reticulum / metabolism Fluorescent Dyes / metabolism Gap Junctions / metabolism* Genes, Dominant Golgi Apparatus / metabolism Hela Cells / metabolism Humans Microscopy, Fluorescence Mutagenesis, Site-Directed Mutation, Missense* Nerve Tissue Proteins / metabolism Neurons / metabolism, ultrastructure PC12 Cells / metabolism Point Mutation* Protein Transport Rats Recombinant Fusion Proteins / physiology Subcellular Fractions / metabolism Transfection |
Grant Support | |
ID/Acronym/Agency:
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NS31224/NS/NINDS NIH HHS |
Chemical | |
Reg. No./Substance:
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0/Connexins; 0/DNA, Complementary; 0/Fluorescent Dyes; 0/Nerve Tissue Proteins; 0/Recombinant Fusion Proteins; 0/connexin 36 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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