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The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation

To cause disease, cholera toxin (CT) is transported from the cell surface to the endoplasmic reticulum (ER) lumen where the catalytic CTA1 subunit retro-translocates to the cytosol to induce pathological water secretion. Two retro-translocon components are the Derlins and ER-associated multi-spannin...

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Autores principales: Bernardi, Kaleena M., Williams, Jeffrey M., Kikkert, Marjolein, van Voorden, Sjaak, Wiertz, Emmanuel J., Ye, Yihong, Tsai, Billy
Formato: Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2801707/
https://www.ncbi.nlm.nih.gov/pubmed/19864457
http://dx.doi.org/10.1091/mbc.E09-07-0586
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author Bernardi, Kaleena M.
Williams, Jeffrey M.
Kikkert, Marjolein
van Voorden, Sjaak
Wiertz, Emmanuel J.
Ye, Yihong
Tsai, Billy
author_facet Bernardi, Kaleena M.
Williams, Jeffrey M.
Kikkert, Marjolein
van Voorden, Sjaak
Wiertz, Emmanuel J.
Ye, Yihong
Tsai, Billy
author_sort Bernardi, Kaleena M.
collection PubMed
description To cause disease, cholera toxin (CT) is transported from the cell surface to the endoplasmic reticulum (ER) lumen where the catalytic CTA1 subunit retro-translocates to the cytosol to induce pathological water secretion. Two retro-translocon components are the Derlins and ER-associated multi-spanning E3 ubiquitin ligases including Hrd1 and gp78. We demonstrated previously that Derlin-1 facilitates CTA1 retro-translocation. However, as CTA1 is neither ubiquitinated on lysines nor at its N-terminus, the role of E3 ligases in toxin retro-translocation is unclear. Here, we show that expression of mutant Hrd1 and gp78 and a mutant E2-conjugating enzyme dedicated to retro-translocation (Ube2g2) decrease CTA1 retro-translocation. Hrd1 knockdown also attenuated toxin retro-translocation. Binding studies demonstrate that Hrd1 and gp78 interact with CT and protein disulfide isomerase, an ER chaperone that unfolds CTA1 to initiate translocation. Moreover, we find that the toxin's association with Hrd1 and gp78 is blocked by dominant-negative Derlin-1, suggesting that CT is targeted initially to Derlin-1 and then transferred to Hrd1 and gp78. These data demonstrate a role of the E3 ubiquitin ligases in CTA1 retro-translocation, implicate a sequence of events experienced by the toxin on the ER membrane, and raise the possibility that ubiquitination is involved in the transport process.
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spelling pubmed-28017072010-03-16 The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation Bernardi, Kaleena M. Williams, Jeffrey M. Kikkert, Marjolein van Voorden, Sjaak Wiertz, Emmanuel J. Ye, Yihong Tsai, Billy Mol Biol Cell Articles To cause disease, cholera toxin (CT) is transported from the cell surface to the endoplasmic reticulum (ER) lumen where the catalytic CTA1 subunit retro-translocates to the cytosol to induce pathological water secretion. Two retro-translocon components are the Derlins and ER-associated multi-spanning E3 ubiquitin ligases including Hrd1 and gp78. We demonstrated previously that Derlin-1 facilitates CTA1 retro-translocation. However, as CTA1 is neither ubiquitinated on lysines nor at its N-terminus, the role of E3 ligases in toxin retro-translocation is unclear. Here, we show that expression of mutant Hrd1 and gp78 and a mutant E2-conjugating enzyme dedicated to retro-translocation (Ube2g2) decrease CTA1 retro-translocation. Hrd1 knockdown also attenuated toxin retro-translocation. Binding studies demonstrate that Hrd1 and gp78 interact with CT and protein disulfide isomerase, an ER chaperone that unfolds CTA1 to initiate translocation. Moreover, we find that the toxin's association with Hrd1 and gp78 is blocked by dominant-negative Derlin-1, suggesting that CT is targeted initially to Derlin-1 and then transferred to Hrd1 and gp78. These data demonstrate a role of the E3 ubiquitin ligases in CTA1 retro-translocation, implicate a sequence of events experienced by the toxin on the ER membrane, and raise the possibility that ubiquitination is involved in the transport process. The American Society for Cell Biology 2010-01-01 /pmc/articles/PMC2801707/ /pubmed/19864457 http://dx.doi.org/10.1091/mbc.E09-07-0586 Text en © 2010 by The American Society for Cell Biology
spellingShingle Articles
Bernardi, Kaleena M.
Williams, Jeffrey M.
Kikkert, Marjolein
van Voorden, Sjaak
Wiertz, Emmanuel J.
Ye, Yihong
Tsai, Billy
The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation
title The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation
title_full The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation
title_fullStr The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation
title_full_unstemmed The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation
title_short The E3 Ubiquitin Ligases Hrd1 and gp78 Bind to and Promote Cholera Toxin Retro-Translocation
title_sort e3 ubiquitin ligases hrd1 and gp78 bind to and promote cholera toxin retro-translocation
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2801707/
https://www.ncbi.nlm.nih.gov/pubmed/19864457
http://dx.doi.org/10.1091/mbc.E09-07-0586
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