Abstract
Cranio-lenticulo-sutural dysplasia (CLSD) is an autosomal recessive syndrome characterized by late-closing fontanels, sutural cataracts, facial dysmorphisms and skeletal defects mapped to chromosome 14q13–q21 (ref. 1). Here we show, using a positional cloning approach, that an F382L amino acid substitution in SEC23A segregates with this syndrome. SEC23A is an essential component of the COPII-coated vesicles that transport secretory proteins from the endoplasmic reticulum to the Golgi complex. Electron microscopy and immunofluorescence show that there is gross dilatation of the endoplasmic reticulum in fibroblasts from individuals affected with CLSD. These cells also exhibit cytoplasmic mislocalization of SEC31. Cell-free vesicle budding assays show that the F382L substitution results in loss of SEC23A function. A phenotype reminiscent of CLSD is observed in zebrafish embryos injected with sec23a-blocking morpholinos. Our observations suggest that disrupted endoplasmic reticulum export of the secretory proteins required for normal morphogenesis accounts for CLSD.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Boyadjiev, S.A. et al. A novel dysmorphic syndrome with open calvarial sutures and sutural cataracts maps to chromosome 14q13–q21. Hum. Genet. 113, 1–9 (2003).
Duden, R. ER-to-Golgi transport: COP I and COP II function (review). Mol. Membr. Biol. 20, 197–207 (2003).
Schekman, R. & Orci, L. Coat proteins and vesicle budding. Science 271, 1526–1533 (1996).
Novick, P., Field, C. & Schekman, R. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway. Cell 21, 205–215 (1980).
Novick, P. & Schekman, R. Secretion and cell-surface growth are blocked in a temperature-sensitive mutant of Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 76, 1858–1862 (1979).
Novick, P. & Schekman, R. Export of major cell surface proteins is blocked in yeast secretory mutants. J. Cell Biol. 96, 541–547 (1983).
Barlowe, C. et al. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum. Cell 77, 895–907 (1994).
Antonny, B. & Schekman, R. ER export: public transportation by the COPII coach. Curr. Opin. Cell Biol. 13, 438–443 (2001).
Miller, E., Antonny, B., Hamamoto, S. & Schekman, R. Cargo selection into COPII vesicles is driven by the Sec24p subunit. EMBO J. 21, 6105–6113 (2002).
Miller, E.A. et al. Multiple cargo binding sites on the COPII subunit Sec24p ensure capture of diverse membrane proteins into transport vesicles. Cell 114, 497–509 (2003).
Nakano, A., Brada, D. & Schekman, R. A membrane glycoprotein, Sec12p, required for protein transport from the endoplasmic reticulum to the Golgi apparatus in yeast. J. Cell Biol. 107, 851–863 (1988).
Barlowe, C. & Schekman, R. SEC12 encodes a guanine-nucleotide-exchange factor essential for transport vesicle budding from the ER. Nature 365, 347–349 (1993).
Aridor, M., Weissman, J., Bannykh, S., Nuoffer, C. & Balch, W.E. Cargo selection by the COPII budding machinery during export from the ER. J. Cell Biol. 141, 61–70 (1998).
Pagano, A. et al. Sec24 proteins and sorting at the endoplasmic reticulum. J. Biol. Chem. 274, 7833–7840 (1999).
Antonny, B., Madden, D., Hamamoto, S., Orci, L. & Schekman, R. Dynamics of the COPII coat with GTP and stable analogues. Nat. Cell Biol. 3, 531–537 (2001).
Bi, X., Corpina, R.A. & Goldberg, J. Structure of the Sec23/24-Sar1 pre-budding complex of the COPII vesicle coat. Nature 419, 271–277 (2002).
Roberts, B., Clucas, C. & Johnstone, I.L. Loss of SEC-23 in Caenorhabditis elegans causes defects in oogenesis, morphogenesis, and extracellular matrix secretion. Mol. Biol. Cell 14, 4414–4426 (2003).
Bottomley, M.J., Batten, M.R., Lumb, R.A. & Bulleid, N.J. Quality control in the endoplasmic reticulum: PDI mediates the ER retention of unassembled procollagen C-propeptides. Curr. Biol. 11, 1114–1118 (2001).
Nakamura, N. et al. Characterization of a cis-Golgi matrix protein, GM130. J. Cell Biol. 131, 1715–1726 (1995).
Kim, J., Hamamoto, S., Ravazzola, M., Orci, L. & Schekman, R. Uncoupled packaging of amyloid precursor protein and presenilin 1 into coat protein complex II vesicles. J. Biol. Chem. 280, 7758–7768 (2005).
Jones, B. et al. Mutations in a Sar1 GTPase of COPII vesicles are associated with lipid absorption disorders. Nat. Genet. 34, 29–31 (2003).
Tiller, G.E. et al. A recurrent RNA-splicing mutation in the SEDL gene causes X-linked spondyloepiphyseal dysplasia tarda. Am. J. Hum. Genet. 68, 1398–1407 (2001).
Zhang, B. et al. Bleeding due to disruption of a cargo-specific ER-to-Golgi transport complex. Nat. Genet. 34, 220–225 (2003).
Fisher, L.W., Lindner, W., Young, M.F. & Termine, J.D. Synthetic peptide antisera: their production and use in the cloning of matrix proteins. Connect. Tissue. Res. 21, 43–48 (1989).
Matsuoka, K. & Schekman, R. The use of liposomes to study COPII- and COPI-coated vesicle formation and membrane protein sorting. Methods 20, 417–428 (2000).
Wilson, R. et al. The translocation, folding, assembly and redox-dependent degradation of secretory and membrane proteins in semi-permeabilized mammalian cells. Biochem. J. 307, 679–687 (1995).
Ben, J., Jabs, E.W. & Chong, S.S. Genomic, cDNA and embryonic expression analysis of zebrafish IRF6, the gene mutated in the human oral clefting disorders Van der Woude and popliteal pterygium syndromes. Gene Expr. Patterns 5, 629–638 (2005).
Acknowledgements
We thank all members of the family that participated in this project; C. Machamer and A. Hubbard for discussions and help with Immunofluorescence; J. Mendell for help with expression vectors, D. Murphy and C. Cooke for assistance with electron microscopy; A. Fischer for help with cell culture; J. Kim and B. Kleizen for assistance with in vitro assays; the Pole Facultaire de Microscopie Ultrastructurale (PFMU) at the University of Geneva Medical School for access to electron microscopy equipment and L. Liu for assistance with zebrafish imaging. This work was supported by grants from the National Institute of Dental and Craniofacial Research–US National Institutes of Health (DE16342 and DE00462 to S.A.B.) and the Swiss National Science Foundation (to L.O.); J.C.F. is supported by a fellowship from the Miller Institute for Basic Research; R.S. is supported by funds from the Howard Hughes Medical Institute and S.S.C. is supported by grants from the NUS (R-178-000-080-112 and R-178-000-104-112).
Author information
Authors and Affiliations
Contributions
This study was initiated by S.A.B., who interpreted clinical and radiologic data, designed and performed experiments, and wrote the manuscript with contributions from J.C.F., R.S., L.O. and S.S.C.; W.E. performed the clinical assessment and provided fibroblast cell lines for these studies; S.A.B., D.J.H. and G.Z. performed the cloning experiments; R.S. and J.C.F. designed, performed and interpreted in vitro liposome-binding and vesicle-formation assays and contributed to immunofluorescence experiments. L.O., M.R. and S.H. performed electron microscopy and immunofluorescence analysis with contributions from C.N. and S.A.B.; L.O. critically interpreted microscopy data and suggested experiments; J.B. and S.S.C. performed the zebrafish expression analysis, the morpholino knockdown experiments and the analysis of the morphants.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Fig. 1
Craniofacial features of CLSD. (PDF 117 kb)
Supplementary Fig. 2
Cranial phenotype of CLSD. (PDF 383 kb)
Supplementary Fig. 3
Skeletal features of CLSD. (PDF 398 kb)
Supplementary Fig. 4
Effectiveness and specificity of translational inhibition of sec23a by antisense morpholinos P and Q. (PDF 79 kb)
Supplementary Table 1
Clnical and radiographic features of four sibs with CLSD as compared with FD, THS, HSS and CCD. (PDF 9 kb)
Supplementary Table 2
Primer and morpholino sequences. (PDF 8 kb)
Rights and permissions
About this article
Cite this article
Boyadjiev, S., Fromme, J., Ben, J. et al. Cranio-lenticulo-sutural dysplasia is caused by a SEC23A mutation leading to abnormal endoplasmic-reticulum-to-Golgi trafficking. Nat Genet 38, 1192–1197 (2006). https://doi.org/10.1038/ng1876
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ng1876
This article is cited by
-
SEC31A may be associated with pituitary hormone deficiency and gonadal dysgenesis
Endocrine (2024)
-
Novel compound heterozygous variants of the SEC23A gene in a Chinese family with cranio-lenticulo-sutural dysplasia based on data from a large cohort of congenital cataract patients
BMC Medical Genomics (2023)
-
The role of vesicle trafficking genes in osteoblast differentiation and function
Scientific Reports (2023)
-
In vitro reconstitution of COPII vesicles from Arabidopsis thaliana suspension-cultured cells
Nature Protocols (2023)
-
ALS/FTD-associated mutation in cyclin F inhibits ER-Golgi trafficking, inducing ER stress, ERAD and Golgi fragmentation
Scientific Reports (2023)