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A membrane protein required for dislocation of misfolded proteins from the ER

Nature volume 429, pages 834–840 (2004)Cite this article

Abstract

After insertion into the endoplasmic reticulum (ER), proteins that fail to fold there are destroyed. Through a process termed dislocation such misfolded proteins arrive in the cytosol, where ubiquitination, deglycosylation and finally proteasomal proteolysis dispense with the unwanted polypeptides. The machinery involved in the extraction of misfolded proteins from the ER is poorly defined. The human cytomegalovirus-encoded glycoproteins US2 and US11 catalyse the dislocation of class I major histocompatibility complex (MHC) products, resulting in their rapid degradation. Here we show that US11 uses its transmembrane domain to recruit class I MHC products to a human homologue of yeast Der1p, a protein essential for the degradation of a subset of misfolded ER proteins. We show that this protein, Derlin-1, is essential for the degradation of class I MHC molecules catalysed by US11, but not by US2. We conclude that Derlin-1 is an important factor for the extraction of certain aberrantly folded proteins from the mammalian ER.

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Figure 1: Identification of US11-associated proteins.
Figure 2: Characterization of Derlin-1.
Figure 3: Association of US11WT and the class I HC with Derlin-1.
Figure 5: Expression of a Derlin-1 dominant-negative construct does not inhibit US2-mediated class I HC degradation, but blocks degradation of the US2 protein itself.
Figure 4: Expression of a Derlin-1 dominant-negative construct impedes US11-mediated class I HC degradation.

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Acknowledgements

We thank B. Kessler and E. Spooner for the preparation of samples and assistance in analysis of mass spectrometry data; R. Tirabassi and K. Ryan for the production of anti-GFP and anti-GRP94 antisera; D. Tortorella for cell lines; and members of the Ploegh laboratory for helpful comments on the manuscript. B.N.L. is a Howard Hughes Medical Institute Predoctoral Fellow. This work was supported by the National Institutes of Health.

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Authors and Affiliations

  1. Department of Pathology, Harvard Medical School, Boston, Massachusetts, 02115, USA

    Brendan N. Lilley & Hidde L. Ploegh

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  1. Brendan N. Lilley
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Correspondence to Hidde L. Ploegh.

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Supplementary information

Supplementary Figure 1

Alignment of human DERtrin proteins. (PDF 66 kb)

Supplementary Figure 2

Analysis of expression levels and localization of DERtrinGFP constructs in US11 and US2 cells. (PDF 226 kb)

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Lilley, B., Ploegh, H. A membrane protein required for dislocation of misfolded proteins from the ER. Nature 429, 834–840 (2004). https://doi.org/10.1038/nature02592

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