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Secretory COPII coat component Sec23a is essential for craniofacial chondrocyte maturation

Nature Genetics volume 38, pages 1198–1203 (2006)Cite this article

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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Figure 1: The crum299 lesion disrupts craniofacial development.
Figure 2: The crum299 mutation is localized in the sec23a gene locus.
Figure 3: Transmission electron microscopy shows sparse ECM deposits and abnormal chondrocyte maturation in crum299 embryos.
Figure 4: The Sec23a mutation disturbs glycosylation and col2a1 expression and triggers the ER stress response.
Figure 5: Knockdown of the sec23b gene produces a phenotype similar to crum299.
Figure 6: Knockdown of the copa gene does not affect chondrocyte maturation.

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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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Author notes

  1. Michael R Lang

    Present address: Department of Biology, University of Washington, Box 351800, Seattle, Washington, 98195, USA

Authors and Affiliations

  1. Department of Medicine, Division of Genetic Medicine, Vanderbilt University Medical Center, Nashville, 37232, Tennessee, USA

    Michael R Lang & Ela W Knapik

  2. Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, 37232, Tennessee, USA

    Michael R Lang, James R Goldenring & Ela W Knapik

  3. Developmental Biology, Institute of Biology I, University of Freiburg, Hauptstrasse 1, Freiburg, D-79104, Germany

    Michael R Lang & Ela W Knapik

  4. Department of Surgery, Vanderbilt University Medical Center, Nashville, 37232, Tennessee, USA

    Lynne A Lapierre & James R Goldenring

  5. Institute of Anatomy and Cell Biology, University of Freiburg, Albertstrasse 17, Freiburg, D-79104, Germany

    Michael Frotscher

  6. Nashville Veterans Administration Medical Center, Nashville, 37212, Tennessee, USA

    James R Goldenring

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Contributions

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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Correspondence to Ela W Knapik.

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The authors declare no competing financial interests.

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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