The VCP/p97 and YOD1 Proteins Have Different Substrate-dependent Activities in Endoplasmic Reticulum-associated Degradation (ERAD)

Sasset, Linda; Petris, Gianluca; Cesaratto, Francesca; Burrone, Óscar R.

doi:10.1074/jbc.m115.656660

Cited by 24 publications

(25 citation statements)

References 72 publications

(74 reference statements)

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“…Indeed, experiments with the proteasome inhibitor MG132 indicated stabilization of the proteins resulting from stalled ribosomes. The same conclusions were obtained using the deubiquitinylase-like protein OTU or the YOD1-C160S, a dominant negative mutant of the ERAD-associated deubiquitinylase YOD1 [35,37,47] and the dominant negative mutant of the p97 ATPase (p97QQ). We also showed that cytosolic proteins produced from 2A* stalled free ribosomes were degraded by the proteasome.…”

Section: Discussionsupporting

confidence: 67%

“…5d). OTU removes ubiquitin from cytosolic poly-ubiquitinylated proteins targeted to degradation, impairing their engagement by the proteasome and therefore causing accumulation [35,36]. Similarly, co-expression with the dominant negative mammalian deubiquitinylase YOD1 (YOD1-C160S) [37] or the dominant negative mutant of the AAA p97 ATPase, which lacks ATPase activity (p97QQ, E305 and E578 mutated into Q) [38], blocked degradation of pr1 from construct 2A*, which accumulated intracellularly further confirming degradation through the ubiquitin-proteasome pathway (Fig.…”

Section: Proteasome Degradation Of Products Of Stalled Ribosomesmentioning

confidence: 99%

“…6b), consistent with active retro-translocation from the ER lumen. Upon coexpression with the dominant negative chaperone BiP/GRP78 (T37G), which binds to newly synthesized protein substrates but is unable to undergo the conformational change upon ATP binding required for protein release, a large accumulation of pr1-2A* was observed, similarly to other ERAD targeted BiP substrates [35,40]. This material was most likely trapped inside the ER, due to its strong interaction with the BiP mutant, as it was not biotinylated by the cytosolic biotin ligase (Fig.…”

Section: Stalling Of Membrane-bound Ribosomes Triggers Eradmentioning

confidence: 99%

“…The coding sequence of BioID2 was previously described [42]. Plasmids expressing cyt-BirA and BiP mutant T37G were previously described [35]. CCHFV-L OTU plasmid was kindly provided by Adolfo García-Sastre, the N-terminal FLAG-tagged human YOD1-C160S plasmid by Christian Schlieker, and the His6-tagged rat p97QQ plasmids by Linda Hendershot.…”

Section: Constructsmentioning

confidence: 99%

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BiP/GRP78 Mediates ERAD Targeting of Proteins Produced by Membrane-Bound Ribosomes Stalled at the STOP-Codon

Cesaratto

Sasset

Myers

et al. 2019

Journal of Molecular Biology

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Translational stalling of ribosome bound to endoplasmic reticulum (ER) membrane requires an accurate clearance of the associated polypeptides, which is not completely understood in mammals. We characterized in mammalian cells the model of ribosomal stalling at the STOP-codon based on proteins tagged at the C-terminus with the picornavirus 2A peptide followed by a termination codon instead of the Proline (2A*). We exploited the 2A* stalling model to characterize the pathway of degradation of ER-targeted polypeptides. We report that the ER chaperone BiP/GRP78 is a new main factor involved. Moreover, degradation of the ER-stalled polypeptides required the activities of the AAA-ATPase VCP/p97, its associated deubiquitinylase YOD1, the ribosome-associated ubiquitin ligase Listerin and the proteasome. In human proteome, we found two human C-terminal amino acid sequences that cause similar stalling at the STOP-codon. Our data suggest that translational stalling at the ER membrane activates protein degradation at the interface of ribosomal-and ERassociated quality control systems.

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Section: Discussionsupporting

confidence: 67%

Section: Proteasome Degradation Of Products Of Stalled Ribosomesmentioning

confidence: 99%

Section: Stalling Of Membrane-bound Ribosomes Triggers Eradmentioning

confidence: 99%

Section: Constructsmentioning

confidence: 99%

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BiP/GRP78 Mediates ERAD Targeting of Proteins Produced by Membrane-Bound Ribosomes Stalled at the STOP-Codon

Cesaratto

Sasset

Myers

et al. 2019

Journal of Molecular Biology

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“…Valosin-containing protein, p97 (VCP/p97, Cdc48 in yeast) [10] is an abundant, conserved ATPase involved in diverse cellular activities, including ERAD, chromatin-associated degradation, mitochondria-associated degradation, and autophagy [11] , [12] , [13] . ERAD requires the retrograde (from ER to cytoplasm) transportation of unfolded proteins, which are ultimately destroyed by the ubiquitin-proteasome system.…”

Section: Introductionmentioning

confidence: 99%

Valosin-Containing Protein/p97 as a Novel Therapeutic Target in Acute Lymphoblastic Leukemia

et al. 2017

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B acute lymphoblastic leukemia (B-ALL) cells are distinctively vulnerable to endoplasmic reticulum (ER) stress. Recently, inhibition of p97 was shown to induce ER stress and subsequently cell death in solid tumors and in multiple myeloma. We investigated the role of a novel, orally available, p97 inhibitor (CB-5083; Cleave Biosciences) in B-ALL. CB-5083 induced a significant reduction in viability in 10 human B-ALL cell lines, harboring the most common fusion-genes involved in pediatric and adult B-ALL, with IC50s ranging from 0.34 to 0.76 μM. Moreover, CB-5083 significantly reduced the colony formation of OP1 and NALM6 cells. Early and strong induction of apoptosis was demonstrated in BALL1 and OP1 cells, together with a robust cleavage of PARP. CB-5083 induced ER stress, as documented through: 1) prominent expression of chaperones (GRP78, GRP94, PDI, DNAJC3, and DNAJB9); 2) increased activation of IRE1-alpha, as demonstrated by the splicing of XBP1; and 3) activation of PERK, which resulted in a significant overexpression of CHOP, and its downstream genes. CB-5083 reduced the viability also in GRP78−/−, GRP94−/−, and XBP1−/− cells, suggesting that none of these proteins alone was strictly required for CB-5083 activity. Moreover, we showed that the absence of XBP1 (XBP1−/−) increased the sensitivity to CB-5083, leading to the hypothesis that XBP1 splicing counteracts the activity of CB-5083, probably mitigating ER stress. Finally, vincristine was synergistic with CB-5083 in both BALL1 and OP1 cells. In summary, the targeting of p97 with CB-5083 is a novel promising therapeutic approach that should be further evaluated in B-ALL.

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Identification of potential diagnostic and prognostic biomarkers for sepsis based on machine learning

Li¹,

Lu²,

Han³

et al. 2023

Computational and Structural Biotechnology Journal

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The VCP/p97 and YOD1 Proteins Have Different Substrate-dependent Activities in Endoplasmic Reticulum-associated Degradation (ERAD)

Cited by 24 publications

References 72 publications

BiP/GRP78 Mediates ERAD Targeting of Proteins Produced by Membrane-Bound Ribosomes Stalled at the STOP-Codon

BiP/GRP78 Mediates ERAD Targeting of Proteins Produced by Membrane-Bound Ribosomes Stalled at the STOP-Codon

Valosin-Containing Protein/p97 as a Novel Therapeutic Target in Acute Lymphoblastic Leukemia

Identification of potential diagnostic and prognostic biomarkers for sepsis based on machine learning

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