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The ubiquitin-selective chaperone CDC-48/p97 links myosin assembly to human myopathy

Nature Cell Biology volume 9, pages 379–390 (2007)Cite this article

An Addendum to this article was published on 01 May 2007

Abstract

Protein degradation in eukaryotes often requires the ubiquitin-selective chaperone p97 for substrate recruitment and ubiquitin-chain assembly. However, the physiological relevance of p97, and its role in developmental processes, remain unclear. Here, we discover an unanticipated function for CDC-48/p97 in myosin assembly and myofibril organization, both in Caenorhabditis elegans and humans. The developmentally regulated assembly of a CDC-48–UFD-2–CHN-1 complex links turnover of the myosin-directed chaperone UNC-45 to functional muscle formation. Our data suggest a similarly conserved pathway regulating myosin assembly in humans. Remarkably, mutations in human p97, known to cause hereditary inclusion-body myopathy, abrogate UNC-45 degradation and result in severely disorganized myofibrils, detrimental towards sarcomeric function. These results identify a key role for CDC-48/p97 in the process of myofibre differentiation and maintenance, which is abolished during pathological conditions leading to protein aggregation and inclusion-body formation in human skeletal muscle.

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Figure 1: UFD-2 binds to the p97 homologues CDC-48.1 and CDC-48.2.
Figure 2: UFD-2 and CHN-1 are not critical for ERAD.
Figure 3: CDC-48 regulates the myosin chaperone UNC-45.
Figure 4: Complex of UFD-2, CHN-1, UNC-45 and CDC-48.1.
Figure 5: Developmental regulation of UNC-45 turnover.
Figure 6: Human p97 is able to compensate loss of CDC-48.1 in vivo.
Figure 7: Stability of human SM UNC-45 depends on UFD2a, p97 and the proteasome.
Figure 8: Myosin assembly regulation is abrogated by the p97R155C mutation.

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Acknowledgements

We are grateful to H. F. Epstein for providing us with Punc-54::unc-45FLAG nematodes and the monoclonal anti-UNC-54 antibody; A. Fire, V. E. Kimonis, Y. Kohara, J. A. Mahoney, S. Mitani, D. Ron, M. Vidal, E. E. Wanker, K. Yamanaka, the Caenorhabditis Genetics Center (funded by the National Institutes of Health (NIH) Center for Research Resources) and the Dana-Farber Cancer Institute and Geneservice Ltd for antibodies, plasmids, cDNAs, and strains. Primary human myoblasts of p97R155C IBMPFD patients were obtained from the Muscle Tissue Culture Collection (MD-NET; service structure S1; 01GM0601), a partner of EuroBioBank (www.eurobiobank.org). We also thank A. Buchberger, C. Eckmann, H. F. Epstein, O. Hobert and S. Jentsch for critical reading of the manuscript. This work was supported by grants from the Deutsche Forschungsgemeinschaft to T.H. (Ho SFB 444/3 and Ho 2541/1-1) and by a grant from the Myositis Association (Washington, DC,) to S.K. and H.L.

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  1. Philipp Christoph Janiesch and Johnny Kim: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre for Molecular Neurobiology (ZMNH), University of Hamburg, Falkenried 94, Hamburg, 20251, Germany

    Philipp Christoph Janiesch, Johnny Kim, Julien Mouysset, Roja Barikbin & Thorsten Hoppe

  2. Friedrich-Baur-Institute and Department of Neurology, Ludwig Maximilians University, Marchioninistr. 17, Munich, 81377, Germany

    Hanns Lochmüller & Sabine Krause

  3. IFOM-FIRC Institute of Molecular Oncology, Via Adamello 16, Milano, 20139, Italy

    Giuseppe Cassata

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Contributions

P.C.J. and J.K. were responsible for the experimental work with support from J.M., R.B. and S.K. G.C. provided technical assistance and H.L. provided the primary patient myoblasts. T.H. planned the entire project, designed experiments, analysed data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Thorsten Hoppe.

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Janiesch, P., Kim, J., Mouysset, J. et al. The ubiquitin-selective chaperone CDC-48/p97 links myosin assembly to human myopathy. Nat Cell Biol 9, 379–390 (2007). https://doi.org/10.1038/ncb1554

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