Ubiquitin-dependent regulation of COPII coat size and function

Abstract

Packaging of proteins from the endoplasmic reticulum into COPII vesicles is essential for secretion. In cells, most COPII vesicles are approximately 60–80 nm in diameter, yet some must increase their size to accommodate 300–400 nm procollagen fibres or chylomicrons. Impaired COPII function results in collagen deposition defects, cranio-lenticulo-sutural dysplasia, or chylomicron retention disease, but mechanisms to enlarge COPII coats have remained elusive. Here, we identified the ubiquitin ligase CUL3–KLHL12 as a regulator of COPII coat formation. CUL3–KLHL12 catalyses the monoubiquitylation of the COPII-component SEC31 and drives the assembly of large COPII coats. As a result, ubiquitylation by CUL3–KLHL12 is essential for collagen export, yet less important for the transport of small cargo. We conclude that monoubiquitylation controls the size and function of a vesicle coat.

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Figure 1: CUL3 regulates mouse ES cell morphology.
Figure 2: KLHL12 is a substrate adaptor for CUL3 in mouse ES cells.
Figure 3: CUL3–KLHL12 monoubiquitylates SEC31.
Figure 4: CUL3–KLHL12-dependent monoubiquitylation enlarges COPII-structures.
Figure 5: CUL3–KLHL12 promotes collagen export.

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Acknowledgements

We thank B. Schulman for advice and gifts of cDNAs and proteins. We are grateful to J. Schaletzky for critically reading the manuscript and many discussions. We thank the members of the Rape and Schekman labs for advice and suggestions, L. Lim for providing Cul3-shRNAs, C. Glazier for contributions on BTB protein cloning, and A. Fischer and M. Richner for tissue culture support. This work was funded by grants from the Pew Foundation (M.R.), the NIH (NIGMS-RO1, M.R.; NIH Director’s New Innovator Award, M.R.), and the Howard Hughes Medical Institute (R.S.). L.J. was funded by a CIRM predoctoral fellowship; she is a Tang fellow. K.B.P. is an HFSP long term post-doctoral fellow.

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Experiments were designed by L.J., K.B.P., R.S. and M.R.; L.J. performed the mouse ES cell screen, identified KLHL12 and SEC31, and analysed the role of CUL3 in COPII formation in cells and in collagen export in mouse ES cells; K.B.P. analysed collagen export in fibroblasts; K.E.W. analysed COPII formation in cells; C.B. identified inactive KLHL12; A.G. performed electron micrscopy; L.J., K.B.P. and M.R. prepared the manuscript.

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Correspondence to Michael Rape.

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Jin, L., Pahuja, K., Wickliffe, K. et al. Ubiquitin-dependent regulation of COPII coat size and function. Nature 482, 495–500 (2012). https://doi.org/10.1038/nature10822

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