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Derlin-1 is a rhomboid pseudoprotease required for the dislocation of mutant α-1 antitrypsin from the endoplasmic reticulum

Abstract

The degradation of misfolded secretory proteins is ultimately mediated by the ubiquitin-proteasome system in the cytoplasm, therefore endoplasmic reticulum–associated degradation (ERAD) substrates must be dislocated across the ER membrane through a process driven by the AAA ATPase p97/VCP. Derlins recruit p97/VCP and have been proposed to be part of the dislocation machinery. Here we report that Derlins are inactive members of the rhomboid family of intramembrane proteases and bind p97/VCP through C-terminal SHP boxes. Human Derlin-1 harboring mutations within the rhomboid domain stabilized mutant α-1 antitrypsin (NHK) at the cytosolic face of the ER membrane without disrupting the p97/VCP interaction. We propose that substrate interaction and p97/VCP recruitment are separate functions that are essential for dislocation and can be assigned respectively to the rhomboid domain and the C terminus of Derlin-1. These data suggest that intramembrane proteolysis and protein dislocation share unexpected mechanistic features.

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Figure 1: Derlins belong to the rhomboid family.
Figure 2: p97/VCP binding to Derlin-1 through an SHP box is required for efficient dislocation of NHK.
Figure 3: The GxxxG motif in the Derlin-1 rhomboid domain is required for the dislocation of NHK.
Figure 4: The Derlin-1 WR motif is essential for the dislocation of NHK.

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Acknowledgements

We thank M. Pearce, J. Christianson and C. Richter for their insightful comments on the manuscript. We also thank H. Ploegh (Whitehead Institute of Biomedical Research) and W. Lennarz (Stony Brook University) for generously providing reagents. This work was supported by a grant from the National Institute of General Medical Science (NIGMS) to R.R.K. E.J.G. was supported by a US National Institutes of Health (NIH) predoctoral training grant and J.A.O. is the recipient of a National Research Service Award from the NIH.

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E.J.G. and R.R.K. contributed to the design of all of the experiments and wrote the manuscript. E.J.G. did the experiments and analyses in Figure 1a–d,f,g; Figure 24; and Supplementary Figures 1, 2, 4 and 5. The microscopy experiments in Figure 1e and Supplementary Figures 3 and 6 were conducted by J.A.O. All authors contributed to the interpretation and conclusions of the experiments.

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Correspondence to Ron R Kopito.

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Greenblatt, E., Olzmann, J. & Kopito, R. Derlin-1 is a rhomboid pseudoprotease required for the dislocation of mutant α-1 antitrypsin from the endoplasmic reticulum. Nat Struct Mol Biol 18, 1147–1152 (2011). https://doi.org/10.1038/nsmb.2111

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