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Title: Covalently linked HslU hexamers support a probabilistic mechanism that links ATP hydrolysis to protein unfolding and translocation

Journal Article · · Journal of Biological Chemistry
 [1];  [2];  [3];  [4];  [1];  [1];  [1]
  1. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
  2. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Takeda Pharmaceuticals, Cambridge, MA (United States)
  3. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Stony Brook Univ., NY (United States)
  4. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Stanford Univ., CA (United States)
+ Show Author Affiliations

The HslUV proteolytic machine consists of HslV, a double-ring self-compartmentalized peptidase, and one or two AAA+ HslU ring hexamers that hydrolyze ATP to power the unfolding of protein substrates and their translocation into the proteolytic chamber of HslV. Furthermore, we use genetic tethering and disulfide bonding strategies to construct HslU pseudohexamers containing mixtures of ATPase active and inactive subunits at defined positions in the hexameric ring. Genetic tethering impairs HslV binding and degradation, even for pseudohexamers with six active subunits, but disulfide-linked pseudohexamers do not have these defects, indicating that the peptide tether interferes with HslV interactions. Importantly, pseudohexamers containing different patterns of hydrolytically active and inactive subunits retain the ability to unfold protein substrates and/or collaborate with HslV in their degradation, supporting a model in which ATP hydrolysis and linked mechanical function in the HslU ring operate by a probabilistic mechanism.

Research Organization:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Organization:
National Inst. of Health
Grant/Contract Number:
AI-16892
OSTI ID:
1430301
Journal Information:
Journal of Biological Chemistry, Vol. 292, Issue 14; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 9 works
Citation information provided by
Web of Science

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Cited By (3)

Proteolysis mediated by the membrane‐integrated ATP‐dependent protease FtsH has a unique nonlinear dependence on ATP hydrolysis rates journal May 2019
Structural insights into ATP hydrolysis by the MoxR ATPase RavA and the LdcI-RavA cage-like complex journal January 2020
New tools for automated high-resolution cryo-EM structure determination in RELION-3 journal November 2018

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59 BASIC BIOLOGICAL SCIENCES
ATP-dependent protease
ATPases associated with diverse cellular activities (AAA)
crystal structure
protein engineering
protein turnover
AAA+ protease
mixed hexameric rings
protein unfolding
protein degradation
HslUV