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Missense mutations abolishing DNA binding of the osteoblast-specific transcription factor OSF2/CBFA1 in cleidocranial dysplasia

Abstract

Cleidocranial dysplasia (CCD) is an autosomal dominant disorder characterized by hypoplastic or absent clavicles, large fontanelles, dental anomalies and delayed skeletal development1. The phenotype is suggestive of a generalized defect in ossification and is one of the most common skeletal dysplasias not associated with disproportionate stature. To date, no genetic determinants of ossficiation have been identified. CCD has been mapped to chromosome 6p212,3, where CBFA1, a gene encoding OSF2/CBFA1, a transcriptional activator of osteoblast differentiation, has been localized4,5. Here, we describe two de novo missense mutations, Met175Arg and Ser191Asn, in the OSF2/CBFA1 gene in two patients with CCD. These two mutations result in substitution of highly conserved amino acids in the DNA-binding domain. DNA-binding studies with the mutant polypeptides show that these amino acid substitutions abolish the DNA-binding ability of OSF2/CBFA1 to its known target sequence. Concurrent studies show that heterozygous nonsense mutations in OSF2/CBFA1 also result in CCD6, while mice homozygous for the osf2/cbfa1 null allele exhibit a more severe lethal phenotype7. Thus, these results together suggest that CCD is produced by haploinsufficiency of OSF2/CBFA1 and provide direct genetic evidence that the phenotype is secondary to an alteration of osteoblast differentiation.

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References

  1. Jones, K.L. Smith's recognizable patterns of human malformation (W.B. Saunders Company, Philadelphia, 1997).

  2. Gelb, B.D., Cooper, E., Shevell, M. & Desnick, R.J. Genetic mapping of the cleidocranial dysplasia (CCD) locus on chromosome band 6p21 to include a microdeletion. Am. J. Med. Genet. 58, 200–205 (1995).

    Article  CAS  Google Scholar 

  3. Mundlos, S. et al. Genetic mapping of cleidocranial dysplasia and evidence of a microdeletion in one family. Hum. Mol. Genet. 4, 71–75 (1995).

    Article  CAS  Google Scholar 

  4. Levanon, D. et al. AML1, AML2, and AML3, the human members of the runt domain gene-family: cDNA structure, expression, and chromosomal localization. Genomics 23, 425–32 (1994).

    Article  CAS  Google Scholar 

  5. Ducy, P., Zhang, R., Geoffrey, V., Ridall, A.L. & Karsenty, G. Osf 2 Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89, 747–754 (1997).

    Article  CAS  Google Scholar 

  6. Mundlos, S. et al. Mutations involving the transcription factor CBFA1 cause cleidocranial dysplasia. Cell 89, 773–779 (1997).

    Article  CAS  Google Scholar 

  7. Otto, F. et al. CBFA1, a candidate gene for the cleidocranial dysplasia syndrome, is essential for osteoblast formation and bone development. Cell 89, 765–771 (1997).

    Article  CAS  Google Scholar 

  8. Gergen, J.P. & Wieschaus, E.F. The localized requirements for a gene affecting segmentation in Drosophila: analysis of larvae mosaic for runt. Dev. Biol. 109, 321–335 (1985).

    Article  CAS  Google Scholar 

  9. Kania, M.A., Bonner, A.S., Duffy, J.B. & Gergen, J.P., The Drosophila segmentation gene runt encodes a novel nuclear regulatory protein that is also expressed in the developing nervous system. Genes Dev. 4, 1701–1713 (1990).

    Google Scholar 

  10. Tsai, C. & Gergen, J.P. Gap gene properties of the pair-rule gene runt during Drosophila segmentation. Development. 120 1671–1683 (1994).

    Google Scholar 

  11. Kagoshima, H. et al. The Runt domain identifies a new family of heteromeric transcriptional regulators. Trends Genet. 9, 338–341 (1993).

    Article  CAS  Google Scholar 

  12. Okuda, T. van Deursen, J., Hiebert, S.W., Grosveld, G. & Downing, J.R. AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis. Cell 84, 321–330 (1996).

    Article  CAS  Google Scholar 

  13. Ahn, M.Y., Bae, S.C., Maruyama, M. & Ito, Y. Comparison of the human genomic structure of the Runt domain-encoding PEBP2/CBFalpha gene family. Gene 168, 279–280 (1996).

    Article  CAS  Google Scholar 

  14. loannou, P.A. et al. A new bacteriophage P1-derived vector for the propagation of large human DNA fragments. Nat. Genet. 6, 84–89 (1994).

    Article  Google Scholar 

  15. Ducy, P. & Karsenty, G. Two distinct osteoblast-specific cis-acting elements control expression of a mouse osteocalcin gene. Mol. Cell. Biol. 15, 1858–1869 (1995).

    Article  CAS  Google Scholar 

  16. Kingsley, D.M. et al. The mouse short ear skeletal morphogenesis locus is associated with defects in a bone morphogenetic member of the TGF beta superfamily. Cell 171, 399–410 (1992).

    Article  Google Scholar 

  17. Kingsley, D.M. What do BMPs do in mammals?Clues from the mouse short-ear mutation. Trends Genet. 10, 16–21 (1994).

    Article  CAS  Google Scholar 

  18. Storm, E.E. et al. Limb alterations in brachypodism brachypodism mice due to mutations in a new member of the TGFβ-superfamily. Nature 368, 639–643 (1994).

    Article  CAS  Google Scholar 

  19. Francomano, C.A., Mclntosh, I. & Wilkin, D.J. Bone dysplasias in man* molecular insights. Curr. Opin. Genet Dev. 6, 301–308 (1996).

    Article  CAS  Google Scholar 

  20. Rousseau, F. et al. Mutations of the fibroblast growth factor receptor-3 gene in achondroplasia. Horm. Res. 45, 108–110 (1996).

    Article  CAS  Google Scholar 

  21. Zakany, J. & Duboule, D. Synpolydactyly in mice with a targeted deficiency in the HoxD complex. Nature 384, 69–71 (1996).

    Article  CAS  Google Scholar 

  22. Muragaki, Y., Mundlos, S., Upton, J. & Olsen, B.R. Altered growth and branching patterns in synpolydactyly caused by mutations in HOXD13. Science 272, 548–551 (1996).

    Article  CAS  Google Scholar 

  23. Wagner, T. et al. Autosomal sex reversal and campomelic dysplasia are caused by mutations in and around the SRY-related gene SOX9. Cell 79, 1111–1120 (1994).

    Article  CAS  Google Scholar 

  24. Foster, J.W. et al. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature 372, 525–530 (1994).

    Article  CAS  Google Scholar 

  25. Karaplis, A.C. et al. Lethal skeletal dysplasia from targeted disruption of the parathyroid hormone-related peptide gene. Genes Dev. 8, 277–289 (1994).

    Article  CAS  Google Scholar 

  26. Grigoriadis, A.E. et al. c-Fos: a key regulator of osteoclast-macrophage lineage determination and bone remodeling. Science 266, 443–448 (1994).

    Article  CAS  Google Scholar 

  27. Ausubel, F.M. et al. Current Protocols in Molecular Biology John Wiley & Sons, New York, (1994).

    Google Scholar 

  28. Innis, M.A., Gelfand, D.H., Sninsky, J.J. & White, T.J. PCR Protocols, a Guide to Methods and Applications. Academic. Press, San Diego, CA, (1990).

    Google Scholar 

  29. Baldini, A. & Lindsay, E.A. Mapping human YAC clones by fluorescence in situ hybridization using Alu-PCR from single yeast colonies. in In Situ Hybridization Protocols, Vol 33 (ed. Choo, K.H.A.) 75–85 Human Press, Clifton, NJ, (1994).

    Chapter  Google Scholar 

  30. DeMarchi, J.M., Richards, C.S., Fenwick, R.G., Pace, R. & Beaudet, A.L. A robotics-assisted procedure for large scale cystic fibrosis mutation analysis. Hum. Mutat. 4, 281–290 (1994).

    Article  CAS  Google Scholar 

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Correspondence to Brendan Lee.

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Lee, B., Thirunavukkarasu, K., Zhou, L. et al. Missense mutations abolishing DNA binding of the osteoblast-specific transcription factor OSF2/CBFA1 in cleidocranial dysplasia. Nat Genet 16, 307–310 (1997). https://doi.org/10.1038/ng0797-307

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