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Protein dislocation from the ER requires polyubiquitination and the AAA-ATPase Cdc48

Nature Cell Biology volume 4, pages 134–139 (2002)Cite this article

Abstract

Endoplasmic reticulum (ER)-associated protein degradation by the ubiquitin–proteasome system requires the dislocation of substrates from the ER into the cytosol. It has been speculated that a functional ubiquitin proteasome pathway is not only essential for proteolysis, but also for the preceding export step. Here, we show that short ubiquitin chains synthesized on proteolytic substrates are not sufficient to complete dislocation; the size of the chain seems to be a critical determinant. Moreover, our results suggest that the AAA proteins of the 26S proteasome are not directly involved in substrate export. Instead, a related AAA complex Cdc48, is required for ER-associated protein degradation upstream of the proteasome.

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Figure 1: Loss of polyubiquitination at the ER membrane blocks retrograde transport of a soluble ER degradation substrate.
Figure 2: The AAA-ATPase Rpt4 is not required for the dislocation of CPY*.
Figure 3: Ubiquitination of substrates of low molecular weight is not sufficient to complete dislocation of CPY*.
Figure 4: Cdc48, Ufd1 and Npl4 are required for efficient degradation of ER proteins.
Figure 5: Polyubiquitinated CPY* is membrane-associated in ufd1-1 cells, but soluble in proteasomal mutants.

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Acknowledgements

The authors thank E.S. Johnson, P. Silver, K.-U. Fröhlich, and T.A. Rapoport for the generous gifts of yeast strains and plasmids. We thank the Sommer group for helpful comments on the manuscript. This work was partially supported by grants from the Deutsche Forschungs Gemeinschaft and the Deutsch-Israelische Projektkooperation (DIP) to T.S. and D.H.W. The Marie Curie Training and Mobility of Researchers program of the European Community and the Helmholtz society provided fellowships for E.J.

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  1. Javier Bordallo

    Present address: Area de Farmacologia, Departmento de Medicina, C/ Julian Claveria s/n, E-33006, Oviedo, Spain

Authors and Affiliations

  1. Max-Delbrück Center for Molecular Medicine, Robert-Rössle Strasse 10, Berlin, 13092, Germany

    Ernst Jarosch, Corinna Volkwein & Thomas Sommer

  2. University of Stuttgart, Institute for Biochemistry, Pfaffenwaldring 55, Stuttgart, 70569, Germany

    Christof Taxis, Javier Bordallo & Dieter H. Wolf

  3. Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue., Boston, 02115, Massachusetts, USA

    Daniel Finley

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Correspondence to Thomas Sommer.

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Jarosch, E., Taxis, C., Volkwein, C. et al. Protein dislocation from the ER requires polyubiquitination and the AAA-ATPase Cdc48. Nat Cell Biol 4, 134–139 (2002). https://doi.org/10.1038/ncb746

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