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 journalArticleResearchpeer-review

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

    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

    PY - 2010/6/2

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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.

    U2 - 10.1091/mbc.E09-08-0743

    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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    ER -