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Multiple pathways in the trafficking and assembly of connexin 26, 32 and 43 into gap junction intercellular communication channels

Martin, P, Blundell, G., Ahmad, S., Errington, Rachel Jane and Evans, William Howard 2001. Multiple pathways in the trafficking and assembly of connexin 26, 32 and 43 into gap junction intercellular communication channels. Journal of Cell Science 114 (21) , pp. 3845-3855.

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Abstract

The assembly of gap junctions was investigated in mammalian cells expressing connexin (Cx) 26, 32 and 43 fused to green, yellow or cyan fluorescent proteins (GFP, YFP, CFP). Targeting of Cx32-CFP and 43-GFP to gap junctions and gap junctional communication was inhibited in cells treated with Brefeldin A, a drug that disassembles the Golgi. However gap junctions constructed of Cx26-GFP were only minimally affected by Brefeldin A. Nocodazole, a microtubule disruptor, had little effect on the assembly of Cx43-GFP gap junctions, but perturbed assembly of Cx26-GFP gap junctions. Co-expression of Cx26-YFP and Cx32-CFP in cells treated with Brefeldin A resulted in assembly of gap junctions constructed of Cx26-YFP. Two amino acids that distinguish Cx26 from Cx32 in transmembrane domains were mutated in Cx32 to investigate underlying mechanisms determining trafficking routes to gap junctions. One mutation, Cx32I28L, conferred on it partial Cx26-like trafficking properties as well the post-translational membrane insertion characteristics of Cx26, suggesting that a key determinant regulating trafficking was present in the first transmembrane domain. The results provide a protein trafficking basis for specifying and regulating connexin composition of gap junctions and thus selectivity of intercellular signaling, with Cx32 and 43 trafficking through the secretory pathway and Cx26 also following an alternative pathway.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Subjects: R Medicine > R Medicine (General)
Uncontrolled Keywords: Animals, Biological Transport, Brefeldin A/pharmacology, COS Cells, Cercopithecus aethiops, Connexin 43/genetics, Connexin 43/metabolism, Connexins/genetics, Connexins/metabolism, Cytoskeleton/drug effects, Cytoskeleton/metabolism, Endoplasmic Reticulum/metabolism, Gap Junctions/metabolism, HeLa Cells, Humans, Intracellular Membranes/metabolism, Nocodazole/pharmacology, Recombinant Fusion Proteins/genetics, Recombinant Fusion Proteins/metabolism, Signal Transduction
Publisher: The Company of Biologists Ltd
ISSN: 0021-9533
Last Modified: 18 Aug 2019 00:11
URI: http://orca.cf.ac.uk/id/eprint/66902

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