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

access $199.00 per year only $3.90 per issue Learn more Buy this article * Purchase on SpringerLink * Instant access to full article PDF Buy now Prices may be subject to local taxes which

are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS

MECHANISMS OF COPII COAT ASSEMBLY AND CARGO RECOGNITION IN THE SECRETORY PATHWAY Article 25 March 2025 STRUCTURE OF THE COMPLETE, MEMBRANE-ASSEMBLED COPII COAT REVEALS A COMPLEX INTERACTION

NETWORK Article Open access 01 April 2021 WFS1 FUNCTIONS IN ER EXPORT OF VESICULAR CARGO PROTEINS IN PANCREATIC Β-CELLS Article Open access 30 November 2021 REFERENCES * Leitinger, B.

Transmembrane collagen receptors. _Annu. Rev. Cell Dev. Biol._ 27, 265–290 (2011) Article  CAS  Google Scholar  * Wickström, S. A., Radovanac, K. & Fassler, R. Genetic analyses of

integrin signaling. _Cold Spring Harb. Perspect. Biol._ 10.1101/cshperspect.a005116 (30 December 2010) * Caswell, P. T., Vadrevu, S. & Norman, J. C. Integrins: masters and slaves of

endocytic transport. _Nature Rev. Mol. Cell Biol._ 10, 843–853 (2009) Article  CAS  Google Scholar  * Stephens, L. E. et al. Deletion of beta 1 integrins in mice results in inner cell mass

failure and peri-implantation lethality. _Genes Dev._ 9, 1883–1895 (1995) Article  CAS  Google Scholar  * Chen, S. S., Fitzgerald, W., Zimmerberg, J., Kleinman, H. K. & Margolis, L.

Cell-cell and cell-extracellular matrix interactions regulate embryonic stem cell differentiation. _Stem Cells_ 25, 553–561 (2007) Article  CAS  Google Scholar  * Lang, M. R., Lapierre, L.

A., Frotscher, M., Goldenring, J. R. & Knapik, E. W. Secretory COPII coat component Sec23a is essential for craniofacial chondrocyte maturation. _Nature Genet._ 38, 1198–1203 (2006)

Article  CAS  Google Scholar  * Townley, A. K. et al. Efficient coupling of Sec23–Sec24 to Sec13–Sec31 drives COPII-dependent collagen secretion and is essential for normal craniofacial

development. _J. Cell Sci._ 121, 3025–3034 (2008) Article  CAS  Google Scholar  * Sarmah, S. et al. Sec24D-dependent transport of extracellular matrix proteins is required for zebrafish

skeletal morphogenesis. _PLoS ONE_ 5, e10367 (2010) Article  ADS  Google Scholar  * Ohisa, S., Inohaya, K., Takano, Y. & Kudo, A. _sec24d_ encoding a component of COPII is essential for

vertebra formation, revealed by the analysis of the medaka mutant, _vbi_. _Dev. Biol._ 342, 85–95 (2010) Article  CAS  Google Scholar  * Boyadjiev, S. A. et al. Cranio-lenticulo-sutural

dysplasia is caused by a _SEC23A_ mutation leading to abnormal endoplasmic-reticulum-to-Golgi trafficking. _Nature Genet._ 38, 1192–1197 (2006) Article  CAS  Google Scholar  * Fromme, J. C.

et al. The genetic basis of a craniofacial disease provides insight into COPII coat assembly. _Dev. Cell_ 13, 623–634 (2007) Article  CAS  Google Scholar  * Jensen, D. & Schekman, R.

COPII-mediated vesicle formation at a glance. _J. Cell Sci._ 124, 1–4 (2011) Article  CAS  Google Scholar  * Stagg, S. M. et al. Structural basis for cargo regulation of COPII coat assembly.

_Cell_ 134, 474–484 (2008) Article  CAS  Google Scholar  * Fath, S., Mancias, J. D., Bi, X. & Goldberg, J. Structure and organization of coat proteins in the COPII cage. _Cell_ 129,

1325–1336 (2007) Article  CAS  Google Scholar  * Fromme, J. C. & Schekman, R. COPII-coated vesicles: flexible enough for large cargo? _Curr. Opin. Cell Biol._ 17, 345–352 (2005) Article

  CAS  Google Scholar  * Saito, K. et al. TANGO1 facilitates cargo loading at endoplasmic reticulum exit sites. _Cell_ 136, 891–902 (2009) Article  CAS  Google Scholar  * Saito, K. et al.

cTAGE5 mediates collagen secretion through interaction with TANGO1 at endoplasmic reticulum exit sites. _Mol. Biol. Cell_ 22, 2301–2308 (2011) Article  CAS  Google Scholar  * Wilson, D. G.

et al. Global defects in collagen secretion in a _Mia3_/_TANGO1_ knockout mouse. _J. Cell Biol._ 193, 935–951 (2011) Article  CAS  Google Scholar  * Kathiresan, S. et al. Genome-wide

association of early-onset myocardial infarction with single nucleotide polymorphisms and copy number variants. _Nature Genet._ 41, 334–341 (2009); corrigendum. 41, 762 (2009) * Sumara, I.

et al. A Cul3-based E3 ligase removes Aurora B from mitotic chromosomes, regulating mitotic progression and completion of cytokinesis in human cells. _Dev. Cell_ 12, 887–900 (2007) Article 

CAS  Google Scholar  * Shaw, L. M., Rabinovitz, I., Wang, H. H., Toker, A. & Mercurio, A. M. Activation of phosphoinositide 3-OH kinase by the α6β4 integrin promotes carcinoma invasion.

_Cell_ 91, 949–960 (1997) Article  CAS  Google Scholar  * Chen, Y. et al. Cullin mediates degradation of RhoA through evolutionarily conserved BTB adaptors to control actin cytoskeleton

structure and cell movement. _Mol. Cell_ 35, 841–855 (2009) Article  CAS  Google Scholar  * Furukawa, M. & Xiong, Y. BTB protein Keap1 targets antioxidant transcription factor Nrf2 for

ubiquitination by the Cullin 3-Roc1 ligase. _Mol. Cell. Biol._ 25, 162–171 (2005) Article  CAS  Google Scholar  * Geyer, R., Wee, S., Anderson, S., Yates, J. & Wolf, D. A. BTB/POZ domain

proteins are putative substrate adaptors for cullin 3 ubiquitin ligases. _Mol. Cell_ 12, 783–790 (2003) Article  CAS  Google Scholar  * Pintard, L. et al. The BTB protein MEL-26 is a

substrate-specific adaptor of the CUL-3 ubiquitin-ligase. _Nature_ 425, 311–316 (2003) Article  CAS  ADS  Google Scholar  * Xu, L. et al. BTB proteins are substrate-specific adaptors in an

SCF-like modular ubiquitin ligase containing CUL-3. _Nature_ 425, 316–321 (2003) Article  CAS  ADS  Google Scholar  * Young, R. A. Control of the embryonic stem cell state. _Cell_ 144,

940–954 (2011) Article  CAS  Google Scholar  * Hughes, H. et al. Organisation of human ER-exit sites: requirements for the localisation of Sec16 to transitional ER. _J. Cell Sci._ 122,

2924–2934 (2009) Article  CAS  Google Scholar  * Kim, W. et al. Systematic and quantitative assessment of the ubiquitin-modified proteome. _Mol. Cell_ 44, 325–340 (2011) Article  CAS  Google

Scholar  * Emanuele, M. J. et al. Global identification of modular cullin-RING ligase substrates. _Cell_ 147, 459–474 (2011) Article  CAS  Google Scholar  * Stephens, D. J. & Pepperkok,

R. Imaging of procollagen transport reveals COPI-dependent cargo sorting during ER-to-Golgi transport in mammalian cells. _J. Cell Sci._ 115, 1149–1160 (2002) CAS  Google Scholar  * Zhu, W.

et al. Bcl-2 mutants with restricted subcellular location reveal spatially distinct pathways for apoptosis in different cell types. _EMBO J._ 15, 4130–4141 (1996) Article  CAS  Google

Scholar  * Singer, J. D., Gurian-West, M., Clurman, B. & Roberts, J. M. Cullin-3 targets cyclin E for ubiquitination and controls S phase in mammalian cells. _Genes Dev._ 13, 2375–2387

(1999) Article  CAS  Google Scholar  * Uchida, K. et al. Identification of specific autoantigens in Sjogren’s syndrome by SEREX. _Immunology_ 116, 53–63 (2005) Article  CAS  Google Scholar 

* Angers, S. et al. The KLHL12–Cullin-3 ubiquitin ligase negatively regulates the Wnt–β-catenin pathway by targeting Dishevelled for degradation. _Nature Cell Biol._ 8, 348–357 (2006)

Article  CAS  Google Scholar  * Schäfer, M. & Werner, S. Cancer as an overhealing wound: an old hypothesis revisited. _Nature Rev. Mol. Cell Biol._ 9, 628–638 (2008) Article  Google

Scholar  * Nguyen, D. X., Bos, P. D. & Massague, J. Metastasis: from dissemination to organ-specific colonization. _Nature Rev. Cancer_ 9, 274–284 (2009) Article  CAS  Google Scholar  *

Zhang, X. H. et al. Latent bone metastasis in breast cancer tied to Src-dependent survival signals. _Cancer Cell_ 16, 67–78 (2009) Article  CAS  Google Scholar  * Geddis, A. E. &

Prockop, D. J. Expression of human _COL1A1_ gene in stably transfected HT1080 cells: the production of a thermostable homotrimer of type I collagen in a recombinant system. _Matrix_ 13,

399–405 (1993) Article  CAS  Google Scholar  * Williamson, A. et al. Identification of a physiological E2 module for the human anaphase-promoting complex. _Proc. Natl Acad. Sci. USA_ 106,

18213–18218 (2009) Article  CAS  ADS  Google Scholar  Download references 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. AUTHOR INFORMATION Author notes * Lingyan Jin and Kanika Bajaj Pahuja: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Department of

Molecular and Cell Biology, University of California at Berkeley, California, 94720, USA Lingyan Jin, Kanika Bajaj Pahuja, Katherine E. Wickliffe, Amita Gorur, Christine Baumgärtel, Randy

Schekman & Michael Rape * Howard Hughes Medical Institute, University of California at Berkeley, California, 94720, USA Kanika Bajaj Pahuja, Amita Gorur & Randy Schekman Authors *

Lingyan Jin View author publications You can also search for this author inPubMed Google Scholar * Kanika Bajaj Pahuja View author publications You can also search for this author inPubMed 

Google Scholar * Katherine E. Wickliffe View author publications You can also search for this author inPubMed Google Scholar * Amita Gorur View author publications You can also search for

this author inPubMed Google Scholar * Christine Baumgärtel View author publications You can also search for this author inPubMed Google Scholar * Randy Schekman View author publications You

can also search for this author inPubMed Google Scholar * Michael Rape View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS 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. CORRESPONDING AUTHOR Correspondence to Michael Rape. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests.

SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION This file contains Supplementary Figures 1-8 with legends, Supplementary Methods and Supplementary Tables 1-2. (PDF 5604 kb) POWERPOINT

SLIDES POWERPOINT SLIDE FOR FIG. 1 POWERPOINT SLIDE FOR FIG. 2 POWERPOINT SLIDE FOR FIG. 3 POWERPOINT SLIDE FOR FIG. 4 POWERPOINT SLIDE FOR FIG. 5 RIGHTS AND PERMISSIONS Reprints and

permissions ABOUT THIS ARTICLE CITE THIS ARTICLE 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 Download citation * Received: 03 August 2011 * Accepted: 03 January 2012 * Published: 22 February 2012 * Issue Date: 23 February 2012 * DOI:

https://doi.org/10.1038/nature10822 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently

available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative