Targeted substrate loop insertion by VCP/p97 during PP1 complex disassembly.
Adaptor Proteins, Signal Transducing
/ metabolism
Animals
Binding Sites
/ genetics
Catalytic Domain
/ genetics
Cell Line
Fluorescence Resonance Energy Transfer
HEK293 Cells
Humans
Protein Binding
/ genetics
Protein Phosphatase 1
/ metabolism
Protein Unfolding
Sf9 Cells
Valosin Containing Protein
/ metabolism
Journal
Nature structural & molecular biology
ISSN: 1545-9985
Titre abrégé: Nat Struct Mol Biol
Pays: United States
ID NLM: 101186374
Informations de publication
Date de publication:
12 2021
12 2021
Historique:
received:
22
06
2021
accepted:
07
10
2021
pubmed:
27
11
2021
medline:
7
1
2022
entrez:
26
11
2021
Statut:
ppublish
Résumé
The AAA-ATPase VCP/p97/Cdc48 unfolds proteins by threading them through its central pore, but how substrates are recognized and inserted into the pore in diverse pathways has remained controversial. Here, we show that p97, with its adapter p37, binds an internal recognition site (IRS) within inhibitor-3 (I3) and then threads a peptide loop into its channel to strip I3 off protein phosphatase-1 (PP1). Of note, the IRS is adjacent to the prime interaction site of I3 to PP1, and IRS mutations block I3 processing both in vitro and in cells. In contrast, amino- and carboxy-terminal regions of I3 are not required, and even circularization of I3 does not prevent I3 processing. This was confirmed by an in vitro Förster resonance energy transfer assay that allowed kinetic analysis of the reaction. Thus, our data uncover how PP1 is released from its inhibitory partner for activation and demonstrate a remarkable plasticity in substrate threading by p97.
Identifiants
pubmed: 34824462
doi: 10.1038/s41594-021-00684-5
pii: 10.1038/s41594-021-00684-5
doi:
Substances chimiques
Adaptor Proteins, Signal Transducing
0
p37 protein, human
0
Protein Phosphatase 1
EC 3.1.3.16
VCP protein, human
EC 3.6.4.6
Valosin Containing Protein
EC 3.6.4.6
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
964-971Informations de copyright
© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.
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