Folding-competent and folding-defective forms of Ricin A Chain have different fates after retrotranslocation from the Endoplasmic Reticulum

S Li, R A Spooner, Stuart C H Allen, C P Guise, G Ladds, T Schnoder, M J Schmitt, J M Lord, R M Roberts

Research output: Contribution to JournalArticle

Abstract

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), 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, 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.
Original languageEnglish
Pages (from-to) 2543–2554
Number of pages11
JournalMolecular Biology of the Cell
Volume21
Issue number15
DOIs
Publication statusPublished - 2 Jun 2010

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Ricin
Endoplasmic Reticulum
Cytosol
Proteasome Endopeptidase Complex
Endoplasmic Reticulum-Associated Degradation
Peptides
Ubiquitin-Protein Ligases
Poisons
Ligases
Ubiquitin
Saccharomyces cerevisiae
Adenosine Triphosphatases
Proteins

Cite this

Li, S ; Spooner, R A ; Allen, Stuart C H ; Guise, C P ; Ladds, G ; Schnoder, T ; Schmitt, M J ; Lord, J M ; Roberts, R M. / Folding-competent and folding-defective forms of Ricin A Chain have different fates after retrotranslocation from the Endoplasmic Reticulum. In: Molecular Biology of the Cell. 2010 ; Vol. 21, No. 15. pp. 2543–2554.
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Li, S, Spooner, RA, Allen, SCH, Guise, CP, Ladds, G, Schnoder, T, Schmitt, MJ, Lord, JM & Roberts, RM 2010, 'Folding-competent and folding-defective forms of Ricin A Chain have different fates after retrotranslocation from the Endoplasmic Reticulum', Molecular Biology of the Cell, vol. 21, no. 15, pp. 2543–2554. https://doi.org/10.1091/mbc.E09-08-0743

Folding-competent and folding-defective forms of Ricin A Chain have different fates after retrotranslocation from the Endoplasmic Reticulum. / Li, S; Spooner, R A; Allen, Stuart C H; Guise, C P; Ladds, G; Schnoder, T; Schmitt, M J; Lord, J M; Roberts, R M.

In: Molecular Biology of the Cell, Vol. 21, No. 15, 02.06.2010, p. 2543–2554.

Research output: Contribution to JournalArticle

TY - JOUR

T1 - Folding-competent and folding-defective forms of Ricin A Chain have different fates after retrotranslocation from the Endoplasmic Reticulum

AU - Li, S

AU - Spooner, R A

AU - Allen, Stuart C H

AU - Guise, C P

AU - Ladds, G

AU - Schnoder, T

AU - Schmitt, M J

AU - Lord, J M

AU - Roberts, R M

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N2 - 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), 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, 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.

AB - 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), 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, 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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DO - 10.1091/mbc.E09-08-0743

M3 - Article

VL - 21

SP - 2543

EP - 2554

JO - Molecular Biology of the Cell

JF - Molecular Biology of the Cell

SN - 1059-1524

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