Abstract
An increasing number of human disorders have been linked to mutations in genes of the secretory pathway. The chemically induced zebrafish crusher variant results in malformed craniofacial skeleton, kinked pectoral fins and a short body length. By positional cloning, we identified a nonsense mutation converting leucine to a stop codon (L402X) in the sec23a gene, an integral component of the COPII complex, which is critical for anterograde protein trafficking between endoplasmic reticulum and Golgi apparatus. Zebrafish crusher mutants develop normally until the onset of craniofacial chondrogenesis. crusher chondrocytes accumulate proteins in a distended endoplasmic reticulum, resulting in severe reduction of cartilage extracellular matrix (ECM) deposits, including type II collagen. We demonstrate that the paralogous gene sec23b is also an essential component of the ECM secretory pathway in chondrocytes. In contrast, knockdown of the COPI complex does not hinder craniofacial morphogenesis. As SEC23A lesions cause the cranio-lenticulo-sutural dysplasia syndrome, crusher provides the first vertebrate model system that links the biology of endoplasmic reticulum to Golgi trafficking with a clinically relevant dysmorphology.
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Acknowledgements
We thank C. Knappmeyer and S. Goetter for excellent zebrafish animal care; B. Joch for electron microscopy; S. Ferrone for LMP7 and HLAII antibodies; W. Driever for sharing the crusher mutant and wild-type zebrafish lines and L. Beihoffer for performing supporting experiments. We are indebted to A. Hatzopoulos, T. Graham, A. George and M. Summers for critically reading the manuscript and S. Wente for discussions. This work has been supported in part by the zebrafish initiative of the Vanderbilt University Academic Venture Capital Fund (E.W.K.).
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M.R.L. conducted all experiments except electron microscopy by M.F. and confocal imaging by L.A.L. The project was designed and orchestrated by E.W.K., who analyzed the data and wrote the paper. All authors discussed the results and commented on the manuscript.
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Supplementary information
Supplementary Fig. 1
Endochondral ossification is aborted in crum299 embryos. (PDF 474 kb)
Supplementary Fig. 2
The Sec23 primary structure has been highly conserved in evolution. (PDF 69 kb)
Supplementary Table 1
Oligonucleotides sequences. (PDF 31 kb)
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Lang, M., Lapierre, L., Frotscher, M. et al. Secretory COPII coat component Sec23a is essential for craniofacial chondrocyte maturation. Nat Genet 38, 1198–1203 (2006). https://doi.org/10.1038/ng1880
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DOI: https://doi.org/10.1038/ng1880
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