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Loss-of-function mutations in the EGF-CFC gene CFC1 are associated with human left-right laterality defects

A Correction to this article was published on 01 December 2000


All vertebrates display a characteristic asymmetry of internal organs with the cardiac apex, stomach and spleen towards the left, and the liver and gall bladder on the right1,2,3. Left-right (L-R) axis abnormalities or laterality defects are common in humans (1 in 8,500 live births). Several genes (such as Nodal, Ebaf and Pitx2) have been implicated in L-R organ positioning in model organisms2,3,4. In humans, relatively few genes have been associated with a small percentage of human situs defects. These include ZIC3 (ref. 5), LEFTB (formerly LEFTY2; ref. 6) and ACVR2B (encoding activin receptor IIB; ref. 7). The EGF-CFC genes8, mouse Cfc1 (encoding the Cryptic protein; ref. 9) and zebrafish one-eyed pinhead (oep; refs 10, 11) are essential for the establishment of the L-R axis12,13. EGF-CFC proteins act as co-factors for Nodal-related signals11, which have also been implicated in L-R axis development4. Here we identify loss-of-function mutations in human CFC1 (encoding the CRYPTIC protein) in patients with heterotaxic phenotypes (randomized organ positioning). The mutant proteins have aberrant cellular localization in transfected cells and are functionally defective in a zebrafish oep-mutant rescue assay. Our findings indicate that the essential role of EGF-CFC genes and Nodal signalling in left-right axis formation is conserved from fish to humans. Moreover, our results support a role for environmental and/or genetic modifiers in determining the ultimate phenotype in humans.

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Figure 1: Genomic structure and sequence alignment of CFC1.
Figure 2: Cellular localization of wild-type and variant CRYPTIC proteins in transfected COS cells.
Figure 3: Activities of CFC1 mutations in zebrafish embryos.

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  1. Kosaki, K. & Casey, B. Genetics of human left-right axis malformations. Cell Dev. Biol. 9, 89 –99 (1998).

    Article  CAS  Google Scholar 

  2. Capdevila, J., Vogan, K.J., Tabin, C. & Izpisua Belmonte, J.C. Mechanisms of left-right determination in vertebrates. Cell 101 , 9–21 (2000).

    Article  CAS  Google Scholar 

  3. Burdine, R. & Schier, A.F. Conserved and divergent mechanisms in left-right axis formation. Genes Dev. 14, 763–776 (2000).

    CAS  Google Scholar 

  4. Schier, A.F. & Shen, M.M. Nodal signalling in vertebrate development. Nature 403, 385– 389 (2000).

    Article  CAS  Google Scholar 

  5. Gebbia, M. et al. X-linked situs abnormalities result from mutations in ZIC3. Nature Genet. 17, 305– 308 (1997).

    Article  CAS  Google Scholar 

  6. Kosaki, K. et al. Characterization and mutation analysis of human LEFTYA and LEFTYB, homologues of murine genes implicated in left-right axis development. Am. J. Hum. Genet. 64, 712–721 (1999).

    Article  CAS  Google Scholar 

  7. Kosaki, R. et al. Left-right axis malformations associated with mutations in ACVR2B, the gene for human activin receptor type IIB. Am. J. Med. Genet. 82, 70–76 ( 1999).

    Article  CAS  Google Scholar 

  8. Shen, M.M. & Schier, A.F. The EGF-CFC gene family in vertebrate development. Trends Genet . 16, 303–309 (2000).

    Article  CAS  Google Scholar 

  9. Shen, M.M., Wang, H. & Leder, P. A differential display strategy identifies Cryptic, a novel EGF-related gene expressed in the axial and lateral mesoderm during mouse gastrulation. Development 124, 429 –442 (1997).

    CAS  Google Scholar 

  10. Zhang, J., Talbot, W.S. & Schier, A.F. Positional cloning identifies zebrafish one-eyed pinhead as a permissive EGF-related ligand required during gastrulation. Cell 92, 241–251 ( 1998).

    Article  CAS  Google Scholar 

  11. Gritsman, K. et al. The EGF-CFC protein one-eyed pinhead is essential for nodal signaling. Cell 97, 121– 137 (1999).

    Article  CAS  Google Scholar 

  12. Gaio, U. et al. A role of the cryptic gene in the correct establishment of the left-right axis. Curr. Biol. 9, 1339– 1342 (1999).

    Article  CAS  Google Scholar 

  13. Yan, Y.-T. et al. Conserved requirement for EGF-CFC genes in vertebrate left-right axis formation. Genes Dev. 13, 2527– 2537 (1999).

    Article  CAS  Google Scholar 

  14. Minchiotti, G. et al. Membrane-anchorage of Cripto protein by glycosylphosphatidylinositol and its distribution during early mouse development. Mech. Dev. 90, 133–142 ( 2000).

    Article  CAS  Google Scholar 

  15. Kane, D.A. et al. The zebrafish epiboly mutants. Development 123, 47–55 (1996).

    CAS  Google Scholar 

  16. Solnica-Krezel, L. et al. Mutations affecting cell fates and cellular rearrangements during gastrulation in zebrafish. Development 123, 67–80 (1996).

    CAS  Google Scholar 

  17. Osada, S.I. et al. Activin/nodal responsiveness and asymmetric expression of a Xenopus nodal-related gene converge on a FAST-regulated module in intron 1. Development 127, 2503– 2514 (2000).

    CAS  Google Scholar 

  18. Meno, C. et al. Mouse lefty2 and zebrafish antivin are feedback inhibitors of nodal signaling during vertebrate gastrulation. Mol. Cell 4, 287–298 (1999).

    Article  CAS  Google Scholar 

  19. Niikawa, N., Kohsaka, S., Mizumoto, M., Hamada, I. & Kajii, T. Familial clustering of situs inversus totalis, and asplenia and polysplenia syndromes. Am. J. Med. Genet. 16, 43–47 ( 1983).

    Article  CAS  Google Scholar 

  20. Saijoh, Y. et al. Left-right asymmetric expression of lefty2 and nodal is induced by a signaling pathway that includes the transcription factor FAST2. Mol. Cell 5, 35–47 ( 2000).

    Article  CAS  Google Scholar 

  21. Collignon, D.B., Varlet, I. & Robertson, E. Relationship between asymmetric nodal expression and the direction of embryonic turning. Nature 381, 155–158 (1996).

    Article  CAS  Google Scholar 

  22. Nomura, M. & Li, E. Smad2 role in mesoderm formation, left-right patterning and craniofacial development. Nature 393, 786–790 (1998).

    Article  CAS  Google Scholar 

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We thank the patients and their families for participation. R.D.B. is supported by a postdoctoral fellowship from the Damon Runyon-Walter Winchell Cancer Research Fund. A.F.S. is a Scholar of the McKnight Endowment Fund for Neuroscience. The authors are supported by grants from the NIH (B.C., A.F.S. and M.M.S.), the US Army Breast Cancer Research Program (M.M.S.) and the Division of Intramural Research, National Human Genome Research Institute, NIH (M.M.).

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Correspondence to Maximilian Muenke.

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Bamford, R., Roessler, E., Burdine, R. et al. Loss-of-function mutations in the EGF-CFC gene CFC1 are associated with human left-right laterality defects. Nat Genet 26, 365–369 (2000).

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