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Cited by 37Ubiquitin and proteasome pathwaysHeat shock proteins researchDNA Repair Mechanisms

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Multiple UBX proteins reduce the ubiquitin threshold of the mammalian p97-UFD1-NPL4 unfoldase

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Fujisawa, Ryo, Polo Rivera, Cristian, Labib, Karim PM·2022·eLife
DOI 10.7554/elife.76763PMC PMC9377798

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Abstract

The p97/Cdc48 ATPase and its ubiquitin receptors Ufd1-Npl4 are essential to unfold ubiquitylated proteins in many areas of eukaryotic cell biology. In yeast, Cdc48-Ufd1-Npl4 is controlled by a quality control mechanism, whereby substrates must be conjugated to at least five ubiquitins. Here, we show that mammalian p97-UFD1-NPL4 is governed by a complex interplay between additional p97 cofactors and the number of conjugated ubiquitins. Using reconstituted assays for the disassembly of ubiquitylated CMG (Cdc45-MCM-GINS) helicase by human p97-UFD1-NPL4, we show that the unfoldase has a high ubiquitin threshold for substrate unfolding, which can be reduced by the UBX proteins UBXN7, FAF1, or FAF2. Our data indicate that the UBX proteins function by binding to p97-UFD1-NPL4 and stabilising productive interactions between UFD1-NPL4 and K48-linked chains of at least five ubiquitins. Stimulation by UBXN7 is dependent upon known ubiquitin-binding motifs, whereas FAF1 and FAF2 use a previously uncharacterised coiled-coil domain to reduce the ubiquitin threshold of p97-UFD1-NPL4. We show that deleting the Ubnx7 and Faf1 genes impairs CMG disassembly during S-phase and mitosis and sensitises cells to reduced ubiquitin ligase activity. These findings indicate that multiple UBX proteins are important for the efficient unfolding of ubiquitylated proteins by p97-UFD1-NPL4 in mammalian cells.

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Metadata

Authors
Fujisawa, R., Polo Rivera, C., & Labib, K. P.
Affiliations
  • Harvard University (US)
  • The MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee
Corresponding author
Labib, Karim PM
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2022-08-03
Volume
11
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.76763
PMC
PMC9377798
OpenAlex
W4289550428
Cited by
37
References
3
Citations by year
2026: 11 · 2025: 4 · 2024: 8 · 2023: 11 · 2022: 3
FWCI
2.12
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Discovery Research Portal (University of Dundee) (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Medical Research Council (MC_UU_12016/13)
  • Cancer Research UK (C578/A24558, C578/A25671)
  • Japan Society for the Promotion of Science (202160572)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
Keywords
FAF1, ubiquitin receptors, K48-linked ubiquitin chains, protein unfolding, coiled-coil domain, ubiquitin ligase, S phase, mitosis, protein quality control
MeSH terms
Valosin Containing Protein, Animals, Humans, Nuclear Proteins, Protein Binding, Ubiquitins, Cell Cycle Proteins, Intracellular Signaling Peptides and Proteins, Adaptor Proteins, Signal Transducing, Apoptosis Regulatory Proteins

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