Multisubunit assembly of an integral plasma membrane channel protein, gap junction connexin43, occurs after exit from the ER

Linda S. Musil; Daniel A. Goodenough

doi:10.1016/0092-8674(93)90728-9

Multisubunit assembly of an integral plasma membrane channel protein, gap junction connexin43, occurs after exit from the ER

Linda S. Musil
, Daniel A. Goodenough

Research output: Contribution to journal › Article › peer-review

431 Scopus citations

Abstract

Connexin43 (Cx43) is an integral plasma membrane protein that forms gap junctions between vertebrate cells. We have used sucrose gradient fractionation and chemical cross-linking to study the first step in gap junction assembly, oligomerization of Cx43 monomers into connexon channels. In contrast with other plasma membrane proteins, multisubunit assembly of Cx43 was specifically and completely blocked when endoplasmic reticulum (ER)-to-Golgi transport was inhibited by 15°C incubation, carbonyl cyanide m-chlorophenylhydrazone, or brefeldin A or in CHO cell mutants with temperature-sensitive defects in secretion. Additional experiments indicated that connexon assembly occurred intracellularly, most likely in the trans-Golgi network. These results describe a post-ER assembly pathway for integral membrane proteins and have implications for the relationship between membrane protein oligomerization and intracellular transport.

Original language	English (US)
Pages (from-to)	1065-1077
Number of pages	13
Journal	Cell
Volume	74
Issue number	6
DOIs	https://doi.org/10.1016/0092-8674(93)90728-9
State	Published - Sep 24 1993
Externally published	Yes

Funding

Experiments with the /d/F and /d/G cell lines were conducted in the laboratory of Monty Krieger (Massachusetts Institute of Technology), and we are very grateful to him for providing these cells and especially to Dr. Abby Fisher for her expertise and advice in their use. We also thank Tom White and Roberto Bruuone for their invaluable help with the Xenopus oocyte experiments and Tom Wileman for his gift of the BFA (originally obtained from Dr. Jennifer Lippincott-Schwartz) used in preliminary experiments. This work was supported in part by grants GM-18974 and EY-02430 to D. A. G. from the National Institutes of Health. L. S. M. is a fellow of the Medical Foundation of Boston.

Funders	Funder number
Author National Institutes of Health National Institutes of Health National Institutes of Health National Institutes of Health The Bev Hartig Huntington's Disease Foundation National Institutes of Health
National Eye Institute and Casey Eye Institute	R01EY002430

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/0092-8674(93)90728-9

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title = "Multisubunit assembly of an integral plasma membrane channel protein, gap junction connexin43, occurs after exit from the ER",

abstract = "Connexin43 (Cx43) is an integral plasma membrane protein that forms gap junctions between vertebrate cells. We have used sucrose gradient fractionation and chemical cross-linking to study the first step in gap junction assembly, oligomerization of Cx43 monomers into connexon channels. In contrast with other plasma membrane proteins, multisubunit assembly of Cx43 was specifically and completely blocked when endoplasmic reticulum (ER)-to-Golgi transport was inhibited by 15°C incubation, carbonyl cyanide m-chlorophenylhydrazone, or brefeldin A or in CHO cell mutants with temperature-sensitive defects in secretion. Additional experiments indicated that connexon assembly occurred intracellularly, most likely in the trans-Golgi network. These results describe a post-ER assembly pathway for integral membrane proteins and have implications for the relationship between membrane protein oligomerization and intracellular transport.",

author = "Musil, \{Linda S.\} and Goodenough, \{Daniel A.\}",

note = "Funding Information: Experiments with the /d/F and /d/G cell lines were conducted in the laboratory of Monty Krieger (Massachusetts Institute of Technology), and we are very grateful to him for providing these cells and especially to Dr. Abby Fisher for her expertise and advice in their use. We also thank Tom White and Roberto Bruuone for their invaluable help with the Xenopus oocyte experiments and Tom Wileman for his gift of the BFA (originally obtained from Dr. Jennifer Lippincott-Schwartz) used in preliminary experiments. This work was supported in part by grants GM-18974 and EY-02430 to D. A. G. from the National Institutes of Health. L. S. M. is a fellow of the Medical Foundation of Boston.",

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AU - Goodenough, Daniel A.

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N2 - Connexin43 (Cx43) is an integral plasma membrane protein that forms gap junctions between vertebrate cells. We have used sucrose gradient fractionation and chemical cross-linking to study the first step in gap junction assembly, oligomerization of Cx43 monomers into connexon channels. In contrast with other plasma membrane proteins, multisubunit assembly of Cx43 was specifically and completely blocked when endoplasmic reticulum (ER)-to-Golgi transport was inhibited by 15°C incubation, carbonyl cyanide m-chlorophenylhydrazone, or brefeldin A or in CHO cell mutants with temperature-sensitive defects in secretion. Additional experiments indicated that connexon assembly occurred intracellularly, most likely in the trans-Golgi network. These results describe a post-ER assembly pathway for integral membrane proteins and have implications for the relationship between membrane protein oligomerization and intracellular transport.

AB - Connexin43 (Cx43) is an integral plasma membrane protein that forms gap junctions between vertebrate cells. We have used sucrose gradient fractionation and chemical cross-linking to study the first step in gap junction assembly, oligomerization of Cx43 monomers into connexon channels. In contrast with other plasma membrane proteins, multisubunit assembly of Cx43 was specifically and completely blocked when endoplasmic reticulum (ER)-to-Golgi transport was inhibited by 15°C incubation, carbonyl cyanide m-chlorophenylhydrazone, or brefeldin A or in CHO cell mutants with temperature-sensitive defects in secretion. Additional experiments indicated that connexon assembly occurred intracellularly, most likely in the trans-Golgi network. These results describe a post-ER assembly pathway for integral membrane proteins and have implications for the relationship between membrane protein oligomerization and intracellular transport.

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Multisubunit assembly of an integral plasma membrane channel protein, gap junction connexin43, occurs after exit from the ER

Abstract

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