Head-to-tail interactions of the coiled-coil domains regulate ClpB activity and cooperation with Hsp70 in protein disaggregation

Research output: Contribution to journalJournal articleResearchpeer-review

  • Marta Carroni
  • Kummer, Eva
  • Yuki Oguchi
  • Petra Wendler
  • Daniel K Clare
  • Irmgard Sinning
  • Jürgen Kopp
  • Axel Mogk
  • Bernd Bukau
  • Helen R Saibil

The hexameric AAA+ chaperone ClpB reactivates aggregated proteins in cooperation with the Hsp70 system. Essential for disaggregation, the ClpB middle domain (MD) is a coiled-coil propeller that binds Hsp70. Although the ClpB subunit structure is known, positioning of the MD in the hexamer and its mechanism of action are unclear. We obtained electron microscopy (EM) structures of the BAP variant of ClpB that binds the protease ClpP, clearly revealing MD density on the surface of the ClpB ring. Mutant analysis and asymmetric reconstructions show that MDs adopt diverse positions in a single ClpB hexamer. Adjacent, horizontally oriented MDs form head-to-tail contacts and repress ClpB activity by preventing Hsp70 interaction. Tilting of the MD breaks this contact, allowing Hsp70 binding, and releasing the contact in adjacent subunits. Our data suggest a wavelike activation of ClpB subunits around the ring.DOI: http://dx.doi.org/10.7554/eLife.02481.001.

Original languageEnglish
Pages (from-to)e02481
Publication statusPublished - 2014
Externally publishedYes

Bibliographical note

Copyright © 2014, Carroni et al.

    Research areas

  • Amino Acid Motifs, Cryoelectron Microscopy, Crystallography, X-Ray, Endopeptidase Clp, Escherichia coli/metabolism, Escherichia coli Proteins/chemistry, HSP70 Heat-Shock Proteins/metabolism, Heat-Shock Proteins/chemistry, Imaging, Three-Dimensional, Molecular Dynamics Simulation, Mutant Proteins/chemistry, Negative Staining, Protein Aggregates, Protein Binding, Protein Structure, Tertiary

ID: 257864968