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Grey-lethal mutation induces severe malignant autosomal recessive osteopetrosis in mouse and human

Nature Medicine volume 9, pages 399–406 (2003)Cite this article

Abstract

The spontaneous mouse grey-lethal (gl) mutation is responsible for a coat color defect and for the development of the most severe autosomal recessive form of osteopetrosis. Using a positional cloning approach, we have mapped and isolated the gl locus from a ∼1.5 cM genetic interval. The gl locus was identified in a bacterial artificial chromosome (BAC) contig by functional genetic complementation in transgenic mice. Genomic sequence analysis revealed that the gl mutation is a deletion resulting in complete loss of function. The unique ∼3 kb wild-type transcript is expressed primarily in osteoclasts and melanocytes as well as in brain, kidney, thymus and spleen. The gl gene is predicted to encode a 338–amino acid type I transmembrane protein that localizes to the intracellular compartment. Mutation in the human GL gene leads to severe recessive osteopetrosis. Our studies show that mouse Gl protein function is absolutely required for osteoclast and melanocyte maturation and function.

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Figure 1: Isolation of the mouse gl gene.
Figure 2: Molecular characterization of the gl gene and mutation.
Figure 3: gl expression pattern.
Figure 4: Expression and immunolocalization of the Gl protein.
Figure 5: Alignment of the predicted amino acid sequences of mouse and human Gl proteins (using ClustalW).
Figure 6: DNA and RNA analysis of a patient with infantile malignant osteopetrosis.

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References

  1. Karsenty, G. The genetic transformation of bone biology. Genes Dev. 13, 3037–3051 (1999).

    Article  CAS  Google Scholar 

  2. Teitelbaum, S.L. Bone resorption by osteoclasts. Science 289, 1504–1508 (2000).

    Article  CAS  Google Scholar 

  3. Lazner, F., Gowen, M., Pavasovic, D. & Kola, I. Osteopetrosis and osteoporosis: two sides of the same coin. Hum. Mol. Genet. 8, 1839–1846 (1999).

    Article  CAS  Google Scholar 

  4. Reeves, J.D., August, C.S., Humbert, J.R. & Weston, W.L. Host defense in infantile osteopetrosis. Pediatrics 64, 202–206 (1979).

    CAS  PubMed  Google Scholar 

  5. Wilson, C.J. & Vellodi, A. Autosomal recessive osteopetrosis: diagnosis, management, and outcome. Arch. Dis. Child 83, 449–452 (2000).

    Article  CAS  Google Scholar 

  6. Gerritsen, E.J. et al. Bone marrow transplantation for autosomal recessive osteopetrosis. A report from the Working Party on Inborn Errors of the European Bone Marrow Transplantation Group. J. Pediatr. 125, 896–902 (1994).

    Article  CAS  Google Scholar 

  7. Eapen, M., Davies, S.M., Ramsay, N.K. & Orchard, P.J. Hematopoietic stem cell transplantation for infantile osteopetrosis. Bone Marrow Transplant. 22, 941–946 (1998).

    Article  CAS  Google Scholar 

  8. Frattini, A. et al. Defects in TCIRG1 subunit of the vacuolar proton pump are responsible for a subset of human autosomal recessive osteopetrosis. Nat. Genet. 25, 343–346 (2000).

    Article  CAS  Google Scholar 

  9. Sobacchi, C. et al. The mutational spectrum of human malignant autosomal recessive osteopetrosis. Hum. Mol. Genet. 10, 1767–1773 (2001).

    Article  CAS  Google Scholar 

  10. Kornak, U. et al. Loss of the ClC-7 chloride channel leads to osteopetrosis in mice and man. Cell 104, 205–215 (2001).

    Article  CAS  Google Scholar 

  11. Tondravi, M.M. et al. Osteopetrosis in mice lacking haematopoietic transcription factor PU.1. Nature 386, 81–84 (1997).

    Article  CAS  Google Scholar 

  12. Iotsova, V. et al. Osteopetrosis in mice lacking NF-κB1 and NF-κB2. Nat. Med. 3, 1285–1289 (1997).

    Article  CAS  Google Scholar 

  13. Yoshida, H. et al. The murine mutation osteopetrosis is in the coding region of the macrophage colony stimulating factor gene. Nature 345, 442–444 (1990).

    Article  CAS  Google Scholar 

  14. Dai, X.M. et al. Targeted disruption of the mouse colony-stimulating factor 1 receptor gene results in osteopetrosis, mononuclear phagocyte deficiency, increased primitive progenitor cell frequencies, and reproductive defects. Blood 99, 111–120 (2002).

    Article  CAS  Google Scholar 

  15. Johnson, R.S., Spiegelman, B.M. & Papaioannou, V. Pleiotropic effects of a null mutation in the c-fos proto-oncogene. Cell 71, 577–586 (1992).

    Article  CAS  Google Scholar 

  16. Wang, Z.Q. et al. Bone and haematopoietic defects in mice lacking c-fos. Nature 360, 741–745 (1992).

    Article  CAS  Google Scholar 

  17. Kong, Y.Y. et al. OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature 397, 315–323 (1999).

    Article  CAS  Google Scholar 

  18. Begg, S.K. et al. Delayed hematopoietic development in osteopetrotic (op/op) mice. J. Exp. Med. 177, 237–242 (1993).

    Article  CAS  Google Scholar 

  19. Thesingh, C.W. & Scherft, J.P. Fusion disability of embryonic osteoclast precursor cells and macrophages in the microphthalmic osteopetrotic mouse. Bone 6, 43–52 (1985).

    Article  CAS  Google Scholar 

  20. Hodgkinson, C.A. et al. Mutations at the mouse microphthalmia locus are associated with defects in a gene encoding a novel basic-helix-loop-helix-zipper protein. Cell 74, 395–404 (1993).

    Article  CAS  Google Scholar 

  21. Soriano, P., Montgomery, C., Geske, R. & Bradley, A. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell 64, 693–702 (1991).

    Article  CAS  Google Scholar 

  22. Lomaga, M.A. et al. TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling. Genes Dev. 13, 1015–1024 (1999).

    Article  CAS  Google Scholar 

  23. Li, Y.P., Chen, W., Liang, Y., Li, E. & Stashenko, P. Atp6i-deficient mice exhibit severe osteopetrosis due to loss of osteoclast-mediated extracellular acidification. Nat. Genet. 23, 447–451 (1999).

    Article  CAS  Google Scholar 

  24. Scimeca, J.C. et al. The gene encoding the mouse homologue of the human osteoclast-specific 116-kDa V-ATPase subunit bears a deletion in osteosclerotic (oc/oc) mutants. Bone 26, 207–213 (2000).

    Article  CAS  Google Scholar 

  25. Gruneberg, H. Grey-lethal, a new mutation in the house mouse. J. Heredity 27, 105–109 (1936).

    Article  Google Scholar 

  26. Walker, D.G. Bone resorption restored in osteopetrotic mice by transplants of normal bone marrow and spleen cells. Science 190, 784–785 (1975).

    Article  CAS  Google Scholar 

  27. Rajapurohitam, V. et al. The mouse osteopetrotic grey-lethal mutation induces a defect in osteoclast maturation/function. Bone 28, 513–523 (2001).

    Article  CAS  Google Scholar 

  28. Vacher, J. & Bernard, H. Genetic localization and transmission of the mouse osteopetrotic grey-lethal mutation. Mamm. Genome 10, 239–243 (1999).

    Article  CAS  Google Scholar 

  29. Chalhoub, N., Benachenhou, N. & Vacher, J. Physical and transcriptional map of the mouse Chromosome 10 proximal region syntenic to human 6q16-q21. Mamm. Genome 12, 887–892 (2001).

    Article  CAS  Google Scholar 

  30. Zhang, Q.H. et al. Cloning and functional analysis of cDNAs with open reading frames for 300 previously undefined genes expressed in CD34+ hematopoietic stem/progenitor cells. Genome Res. 10, 1546–1560 (2000).

    Article  CAS  Google Scholar 

  31. Brown, D., Hirsch, S. & Gluck, S. Localization of a proton-pumping ATPase in rat kidney. J. Clin. Invest. 82, 2114–2126 (1988).

    Article  CAS  Google Scholar 

  32. Perou, C.M. et al. Identification of the murine beige gene by YAC complementation and positional cloning. Nat. Genet. 13, 303–308 (1996).

    Article  CAS  Google Scholar 

  33. Baron, R., Neff, L., Louvard, D. & Courtoy, P.J. Cell-mediated extracellular acidification and bone resorption: evidence for a low pH in resorbing lacunae and localization of a 100-kD lysosomal membrane protein at the osteoclast ruffled border. J. Cell Biol. 101, 2210–2222 (1985).

    Article  CAS  Google Scholar 

  34. Baron, R. et al. Polarized secretion of lysosomal enzymes: co-distribution of cation-independent mannose-6-phosphate receptors and lysosomal enzymes along the osteoclast exocytic pathway. J. Cell Biol. 106, 1863–1872 (1988).

    Article  CAS  Google Scholar 

  35. Blair, H.C., Teitelbaum, S.L., Ghiselli, R. & Gluck, S. Osteoclastic bone resorption by a polarized vacuolar proton pump. Science 245, 855–857 (1989).

    Article  CAS  Google Scholar 

  36. Palokangas, H., Mulari, M. & Vaananen, H.K. Endocytic pathway from the basal plasma membrane to the ruffled border membrane in bone-resorbing osteoclasts. J. Cell Sci. 110, 1767–1780 (1997).

    CAS  PubMed  Google Scholar 

  37. Vacher, J. & Tilghman, S.M. Dominant negative regulation of the mouse α-fetoprotein gene in adult liver. Science 250, 1732–1735 (1990).

    Article  CAS  Google Scholar 

  38. Bennett, D.C., Cooper, P.J. & Hart, I.R. A line of non-tumorigenic mouse melanocytes, syngeneic with the B16 melanoma and requiring a tumour promoter for growth. Int. J. Cancer 39, 414–418 (1987).

    Article  CAS  Google Scholar 

  39. Emerson, J.A., Vacher, J., Cirillo, L.A., Tilghman, S.M. & Tyner, A.L. The zonal expression of α-fetoprotein transgenes in the livers of adult mice. Dev. Dyn. 195, 55–66 (1992).

    Article  CAS  Google Scholar 

  40. Marcinkiewicz, M., Savaria, D. & Marcinkiewicz, J. The pro-protein convertase PC1 is induced in the transected sciatic nerve and is present in cultured Schwann cells: comparison with PC5, furin and PC7, implication in pro-BDNF processing. Mol. Brain Res. 59, 229–246 (1998).

    Article  CAS  Google Scholar 

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Acknowledgements

The authors thank R. McInnes and J. Horsford for melan-a cells; S. Breton for V-ATPase antibodies; D. Bennett for discussion; D. Lohnes and R. Baron for critical reading of the manuscript; U. Ramenghi and families for their participation; J. Marcinkiewicz for advice; and H. Bernard for technical assistance. M.F. has a studentship from the Fonds pour la Formation de Chercheurs et d'Aide à la Recherche (FCAR). This work was supported by a grant from the Canadian Institutes of Health Research and by Hoechst Marion Roussel grant no. R97047 to J.V., partially supported by grant MIUR-FIRB #RBNE019J9W, and is manuscript #71 of the Genoma2000/ITB Project funded by Fondazione Cariplo.

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  1. Institut de Recherches Cliniques de Montréal, Faculté de Médecine de l'Université de Montréal, Montréal, Québec, Canada

    Nader Chalhoub, Nadia Benachenhou, Venkatesh Rajapurohitam, Monica Pata, Mathieu Ferron & Jean Vacher

  2. Istituto di Tecnologie Biomediche Avanzate, Consiglio Nazionale delle Ricerche, Segrate, Italy

    Annalisa Frattini & Anna Villa

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  1. Nader Chalhoub
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Correspondence to Jean Vacher.

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Chalhoub, N., Benachenhou, N., Rajapurohitam, V. et al. Grey-lethal mutation induces severe malignant autosomal recessive osteopetrosis in mouse and human. Nat Med 9, 399–406 (2003). https://doi.org/10.1038/nm842

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