Abstract

Protein aggregates are the hallmark of stressed and ageing cells, and characterize several pathophysiological states1,2. Healthy metazoan cells effectively eliminate intracellular protein aggregates3,4, indicating that efficient disaggregation and/or degradation mechanisms exist. However, metazoans lack the key heat-shock protein disaggregase HSP100 of non-metazoan HSP70-dependent protein disaggregation systems5,6, and the human HSP70 system alone, even with the crucial HSP110 nucleotide exchange factor, has poor disaggregation activity in vitro4,7. This unresolved conundrum is central to protein quality control biology. Here we show that synergic cooperation between complexed J-protein co-chaperones of classes A and B unleashes highly efficient protein disaggregation activity in human and nematode HSP70 systems. Metazoan mixed-class J-protein complexes are transient, involve complementary charged regions conserved in the J-domains and carboxy-terminal domains of each J-protein class, and are flexible with respect to subunit composition. Complex formation allows J-proteins to initiate transient higher order chaperone structures involving HSP70 and interacting nucleotide exchange factors. A network of cooperative class A and B J-protein interactions therefore provides the metazoan HSP70 machinery with powerful, flexible, and finely regulatable disaggregase activity and a further level of regulation crucial for cellular protein quality control.

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Acknowledgements

We thank A. Mogk for critical reading of the manuscript and S. Ungelenk for Hsp26. This work was funded by the Deutsche Forschungsgemeinschaft (SFB1036, BU617/19-1 to B.B.; EXC257, SFB740 to J.K.), Alexander von Humboldt Foundation Postdoctoral Fellowships (to N.B.N. and A.Sz.), National Institutes of Health (the NIGMS, NIA, NIMS), Ellison Medical Foundation and Daniel F. and Ada L. Rice Foundation (to R.I.M.), German Federal Ministry of Education and Research (BMBF) Virtual Liver Network and EU FEP Flagship Programme Human Brain Project (0315749, 604102 to R.C.W.), Klaus Tschira Foundation (to M.B., A.St. and R.C.W.), Sir Henry Wellcome Postdoctoral Fellowship (to F.S.), ETH Zurich and ERC advanced grant Proteomics v3.0 (233226 to R.A.).

Author information

Affiliations

  1. Center for Molecular Biology of the University of Heidelberg (ZMBH), German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, 69120 Heidelberg, Germany

    • Nadinath B. Nillegoda
    • , Anna Szlachcic
    • , Xuechao Gao
    • , D. Lys Guilbride
    • , Rebecca C. Wade
    • , Matthias P. Mayer
    •  & Bernd Bukau
  2. Leibniz-Institute for Molecular Pharmacology (FMP), 13125 Berlin, Germany

    • Janine Kirstein
    • , Kristin Arnsburg
    •  & Annika Scior
  3. Heidelberg Institute for Theoretical Studies (HITS), 69118 Heidelberg, Germany

    • Mykhaylo Berynskyy
    • , Antonia Stank
    •  & Rebecca C. Wade
  4. Heidelberg Graduate School of Mathematical and Computational Methods for the Sciences, Heidelberg University, 69120 Heidelberg, Germany

    • Antonia Stank
  5. Department of Biology, Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland

    • Florian Stengel
    •  & Ruedi Aebersold
  6. Faculty of Science, University of Zurich, 8057 Zurich, Switzerland

    • Ruedi Aebersold
  7. Interdisciplinary Center for Scientific Computing (IWR), Heidelberg University, 69120 Heidelberg, Germany

    • Rebecca C. Wade
  8. Department of Molecular Biosciences, Rice Institute for Biomedical Research, Northwestern University, Evanston, Illinois 60208, USA

    • Richard I. Morimoto

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Contributions

N.B.N. and B.B. conceived the study. N.B.N., J.K., A.Sz., M.B., A.St., F.S., D.L.G., R.C.W., M.P.M. and B.B. designed the experiments. N.B.N., J.K., A.Sz., M.B., A.St., F.S., K.A., X.G. and A.Sc. performed the experiments. N.B.N., J.K., A.Sz., M.B., A.St., F.S., R.A., R.C.W., R.I.M., D.L.G., M.P.M. and B.B. analysed the data. N.B.N., D.L.G., M.P.M. and B.B. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Nadinath B. Nillegoda or Bernd Bukau.

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https://doi.org/10.1038/nature14884

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