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Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum

We report that a toxic polypeptide retaining the potential to refold upon dislocation from the endoplasmic reticulum (ER) to the cytosol (ricin A chain; RTA) and a misfolded version that cannot (termed RTA(Δ)), follow ER-associated degradation (ERAD) pathways in Saccharomyces cerevisiae that substan...

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Autores principales: Li, Shuyu, Spooner, Robert A., Allen, Stuart C. H., Guise, Christopher P., Ladds, Graham, Schnöder, Tina, Schmitt, Manfred J., Lord, J. Michael, Roberts, Lynne M.
Formato: Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2912342/
https://www.ncbi.nlm.nih.gov/pubmed/20519439
http://dx.doi.org/10.1091/mbc.E09-08-0743
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author Li, Shuyu
Spooner, Robert A.
Allen, Stuart C. H.
Guise, Christopher P.
Ladds, Graham
Schnöder, Tina
Schmitt, Manfred J.
Lord, J. Michael
Roberts, Lynne M.
author_facet Li, Shuyu
Spooner, Robert A.
Allen, Stuart C. H.
Guise, Christopher P.
Ladds, Graham
Schnöder, Tina
Schmitt, Manfred J.
Lord, J. Michael
Roberts, Lynne M.
author_sort Li, Shuyu
collection PubMed
description We report that a toxic polypeptide retaining the potential to refold upon dislocation from the endoplasmic reticulum (ER) to the cytosol (ricin A chain; RTA) and a misfolded version that cannot (termed RTA(Δ)), follow ER-associated degradation (ERAD) pathways in Saccharomyces cerevisiae that substantially diverge in the cytosol. Both polypeptides are dislocated in a step mediated by the transmembrane Hrd1p ubiquitin ligase complex and subsequently degraded. Canonical polyubiquitylation is not a prerequisite for this interaction because a catalytically inactive Hrd1p E3 ubiquitin ligase retains the ability to retrotranslocate RTA, and variants lacking one or both endogenous lysyl residues also require the Hrd1p complex. In the case of native RTA, we established that dislocation also depends on other components of the classical ERAD-L pathway as well as an ongoing ER–Golgi transport. However, the dislocation pathways deviate strikingly upon entry into the cytosol. Here, the CDC48 complex is required only for RTA(Δ), although the involvement of individual ATPases (Rpt proteins) in the 19S regulatory particle (RP) of the proteasome, and the 20S catalytic chamber itself, is very different for the two RTA variants. We conclude that cytosolic ERAD components, particularly the proteasome RP, can discriminate between structural features of the same substrate.
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spelling pubmed-29123422010-10-16 Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum Li, Shuyu Spooner, Robert A. Allen, Stuart C. H. Guise, Christopher P. Ladds, Graham Schnöder, Tina Schmitt, Manfred J. Lord, J. Michael Roberts, Lynne M. Mol Biol Cell Articles We report that a toxic polypeptide retaining the potential to refold upon dislocation from the endoplasmic reticulum (ER) to the cytosol (ricin A chain; RTA) and a misfolded version that cannot (termed RTA(Δ)), follow ER-associated degradation (ERAD) pathways in Saccharomyces cerevisiae that substantially diverge in the cytosol. Both polypeptides are dislocated in a step mediated by the transmembrane Hrd1p ubiquitin ligase complex and subsequently degraded. Canonical polyubiquitylation is not a prerequisite for this interaction because a catalytically inactive Hrd1p E3 ubiquitin ligase retains the ability to retrotranslocate RTA, and variants lacking one or both endogenous lysyl residues also require the Hrd1p complex. In the case of native RTA, we established that dislocation also depends on other components of the classical ERAD-L pathway as well as an ongoing ER–Golgi transport. However, the dislocation pathways deviate strikingly upon entry into the cytosol. Here, the CDC48 complex is required only for RTA(Δ), although the involvement of individual ATPases (Rpt proteins) in the 19S regulatory particle (RP) of the proteasome, and the 20S catalytic chamber itself, is very different for the two RTA variants. We conclude that cytosolic ERAD components, particularly the proteasome RP, can discriminate between structural features of the same substrate. The American Society for Cell Biology 2010-08-01 /pmc/articles/PMC2912342/ /pubmed/20519439 http://dx.doi.org/10.1091/mbc.E09-08-0743 Text en © 2010 by The American Society for Cell Biology
spellingShingle Articles
Li, Shuyu
Spooner, Robert A.
Allen, Stuart C. H.
Guise, Christopher P.
Ladds, Graham
Schnöder, Tina
Schmitt, Manfred J.
Lord, J. Michael
Roberts, Lynne M.
Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum
title Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum
title_full Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum
title_fullStr Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum
title_full_unstemmed Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum
title_short Folding-competent and Folding-defective Forms of Ricin A Chain Have Different Fates after Retrotranslocation from the Endoplasmic Reticulum
title_sort folding-competent and folding-defective forms of ricin a chain have different fates after retrotranslocation from the endoplasmic reticulum
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2912342/
https://www.ncbi.nlm.nih.gov/pubmed/20519439
http://dx.doi.org/10.1091/mbc.E09-08-0743
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