The Non-dominant AAA+ Ring in the ClpAP Protease Functions as an Anti-stalling Motor to Accelerate Protein Unfolding and Translocation.
AAA+ proteases
ATP motors
ClpAP
Hsp104/ClpB
optical tweezers
protein degradation
protein translocation
protein unfolding
single-molecule force spectroscopy
translocation pauses
Journal
Cell reports
ISSN: 2211-1247
Titre abrégé: Cell Rep
Pays: United States
ID NLM: 101573691
Informations de publication
Date de publication:
25 02 2020
25 02 2020
Historique:
received:
14
06
2019
revised:
25
11
2019
accepted:
30
01
2020
entrez:
27
2
2020
pubmed:
27
2
2020
medline:
18
3
2021
Statut:
ppublish
Résumé
ATP-powered unfoldases containing D1 and D2 AAA+ rings play important roles in protein homeostasis, but uncertainty about the function of each ring remains. Here we use single-molecule optical tweezers to assay mechanical unfolding and translocation by a variant of the ClpAP protease containing an ATPase-inactive D1 ring. This variant displays substantial mechanical defects in both unfolding and translocation of protein substrates. Notably, when D1 is hydrolytically inactive, ClpAP often stalls for times as long as minutes, and the substrate can back-slip through the enzyme when ATP concentrations are low. The inactive D1 variant also has more difficulty traveling in the N-to-C direction on a polypeptide track than it does moving in a C-to-N direction. These results indicate that D1 normally functions as an auxiliary/regulatory motor to promote uninterrupted enzyme advancement that is fueled largely by the D2 ring.
Identifiants
pubmed: 32101742
pii: S2211-1247(20)30154-6
doi: 10.1016/j.celrep.2020.01.110
pmc: PMC7888974
mid: NIHMS1667320
pii:
doi:
Substances chimiques
Escherichia coli Proteins
0
Adenosine Triphosphate
8L70Q75FXE
ClpA protease, E coli
EC 3.4.21.53
ClpP protease, E coli
EC 3.4.21.92
Endopeptidase Clp
EC 3.4.21.92
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2644-2654.e3Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM049224
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI015706
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Organisme : NIAID NIH HHS
ID : R37 AI015706
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM101988
Pays : United States
Informations de copyright
Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests The authors declare no competing interests.
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