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Cranio-lenticulo-sutural dysplasia is caused by a SEC23A mutation leading to abnormal endoplasmic-reticulum-to-Golgi trafficking

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.

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Figure 1: Craniofacial features of CLSD with depiction of the 1144T→C SEC23A mutation.
Figure 2: Immunofluorescence analysis.
Figure 3: Electron microscopy.
Figure 4: In vitro studies of F382L SEC23A.
Figure 5: Developmental expression of sec23a and loss-of-function phenotype in zebrafish.

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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).

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Authors

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

Correspondence to Simeon A Boyadjiev.

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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)

Supplementary Note (PDF 12 kb)

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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

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