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Methylation sensitive protein binding to an intragenic active X-specific methylated region in theM. robustus HPRT gene

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Somatic Cell and Molecular Genetics

Abstract

An intragenic region of the wallaroo (Macropus robustus) hypoxanthine phosphoribosyltransferase gene which contains three active X-specific methylated cytosines was examined for protein(s) binding.In vitro DNase I footprinting of unmethylated DNA identified three footprints, one of which (footprint I) contained two of the known differentially methylated sites (a HpaII and a HhaI site). Methylation of the footprint I HpaII site only, abolished the formation of several, specific DNA-protein complexes in mobility shift assays. UV cross-linking experiments indicated that polypeptides involved in the methylation sensitive interactions with footprint I had molecular weights ranging from 72 to 48 kDa. Analogous results were obtained with nuclear extracts from both eutherian and metatherian cells, indicating that these proteins are conserved. We suggest that the binding of these proteins to the inactive X may play some role in gene silencing, with the active gene being protected from this effect by methylation of the binding site.

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Literature Cited

  1. Grant, S.G., and Chapman, V.M. (1988).Annu. Rev. Genet. 22:199–233.

    Article  PubMed  CAS  Google Scholar 

  2. Yen, P.H., Patel, P., Chinault, A.C., Mohandas, T., and Shapiro, L.J. (1984).Proc. Natl. Acad. Sci. U.S.A. 81:1759–1763.

    PubMed  CAS  Google Scholar 

  3. Wolf, S.F., Jolly, D.J., Lunnen, K.D., Friedmann, T., and Migeon, B.R. (1984).Proc. Natl. Acad. Sci. U.S.A. 81:2806–2810.

    PubMed  CAS  Google Scholar 

  4. Keith, D.H., Singer-Sam, J., and Riggs, A.D. (1986).Mol. Cell. Biol. 6:4122–4125.

    PubMed  CAS  Google Scholar 

  5. Toniolo, D., Martini G., Migeon, B.R., and Dono, R. (1988).EMBO J. 7:401–406.

    PubMed  CAS  Google Scholar 

  6. Mohandas, T., Sparkes, R.S., and Shapiro, L.J. (1981).Science 211:393–396.

    PubMed  CAS  Google Scholar 

  7. Jones, P.A., Taylor, S.M., Mohandas, T., and Shapiro, L.J. (1982).Proc. Natl. Acad. Sci. U.S.A. 79:1215–1219.

    PubMed  CAS  Google Scholar 

  8. Hors-Cayla, M.C., Heuretz, S., and Frezal, J. (1983).Somatic Cell Mol. Genet. 9:645–657.

    CAS  Google Scholar 

  9. Hornstra, I.K., and Yang, T.P. (1994),Mol. Cell Biol. 14:1419–1430.

    PubMed  CAS  Google Scholar 

  10. Kaslow, D.C., and Migeon, B.R. (1987).Proc. Natl. Acad. Sci. U.S.A. 84:6210–6214.

    PubMed  CAS  Google Scholar 

  11. Loebel, D.A.F., and Johnston, P.G. (1996).Genome Res. 6:114–123.

    PubMed  CAS  Google Scholar 

  12. Bartlett, M.H., Adra, C.N., Park, J., Chapman, V.M., and McBurney, M.W. (1991).Somat. Cell Mol. Genet. 17:35–47.

    Article  PubMed  CAS  Google Scholar 

  13. Lock, L.F., Takagi, N., and Martin, G.R. (1987).Cell 48:39–46.

    Article  PubMed  CAS  Google Scholar 

  14. Singer-Sam, J., Grant, M., LeBon, J.M., Okuyama, K., Chapman, V., Monk, M., and Riggs, A.D. (1990).Mol. Cell Biol. 10:4987–4989.

    PubMed  CAS  Google Scholar 

  15. Grant, M., Zuccotti, M., and Monk, M. (1992).Nature Gen. 2:161–166.

    CAS  Google Scholar 

  16. Stöger, R., Kubicka, P., Liu, C.-G., Kafri, T., Razin, A., Cedar, H., and Barlow, D.P. (1993).Cell 73:61–71.

    PubMed  Google Scholar 

  17. Li, E., Beard, C., and Jaenisch, R. (1993).Nature 366:362–365.

    PubMed  CAS  Google Scholar 

  18. Yen, P.H., Mohandas, T., and Shapiro, L.J. (1986).Somat. Cell Mol. Genet. 12:153–161.

    Article  PubMed  CAS  Google Scholar 

  19. Chong, S., Taylor, K.A., and Piper, A.A. (1995).Biochimica et Biophysica Acta 1264:103–114.

    PubMed  Google Scholar 

  20. Piper, A.A., Bennett, A.M., Noyce, L., Swanton, M.K., and Cooper, D.W. (1993).Somat. Cell Mol. Genet. 19:141–159.

    Article  PubMed  CAS  Google Scholar 

  21. Pearson, W.R., and Lipman, D.J. (1988).Proc. Natl. Acad. Sci. U.S.A. 85:2444–2448.

    PubMed  CAS  Google Scholar 

  22. Prestridge, D.S. (1991).CABIOS 7:203–206.

    PubMed  CAS  Google Scholar 

  23. Thompson, J.D., Higgins, D.G., and Gibson, T.J. (1994).Nucleic Acids Res. 22:4673–4680.

    PubMed  CAS  Google Scholar 

  24. Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York).

    Google Scholar 

  25. Shapiro, D.J., Sharp, P.A., Wahli, W.W., and Keller, M.J. (1988).DNA 7:47–55.

    PubMed  CAS  Google Scholar 

  26. Deryckere, F., and Gannon, F. (1994).Biotechniques 16:405.

    PubMed  CAS  Google Scholar 

  27. Bradford, M.M. (1976).Anal. Biochem. 72:248–254.

    Article  PubMed  CAS  Google Scholar 

  28. Faisst, S., and Meyer, S. (1992).Nucleic Acids Res. 20:3–26.

    PubMed  CAS  Google Scholar 

  29. Labbé, S., Prévost, J., Remondelli, P., Leone, A., and Séguin, C. (1991).Nucleic Acids Res. 19:4225–4231.

    PubMed  Google Scholar 

  30. Augereau, P., and Chambon, P. (1986).EMBO J. 5:1791–1797.

    PubMed  CAS  Google Scholar 

  31. Blackwell, T.K., and Weintraub, H. (1990).Science 250:1104–1110.

    PubMed  CAS  Google Scholar 

  32. Séguin, C., and Hamer, D.H. (1987).Science 235:1383–1387.

    PubMed  Google Scholar 

  33. Inman, R.B., and Schnös, M. (1970).J. Mol. Biol. 49:93–98.

    Article  PubMed  CAS  Google Scholar 

  34. David, J., Gordon, J.S., and Rutter, W.J. (1974).Proc. Natl. Acad. Sci. U.S.A. 71:2808–2812.

    PubMed  CAS  Google Scholar 

  35. Lin, S.Y., and Riggs, A.D. (1972).Proc. Natl. Acad. Sci. U.S.A. 69:2574–2576.

    PubMed  CAS  Google Scholar 

  36. Kumar, G., Sharma, A.K., and Jayaraman, K. (1992).Oncogene 7:1453–1455.

    PubMed  CAS  Google Scholar 

  37. Coulondre, C., Miller, J.H., Farabaugh, P.J., and Gilbert, W. (1978).Nature 274:775–780.

    Article  PubMed  CAS  Google Scholar 

  38. Ehrlich, M., and Ehrlich, K.C. (1993). InDNA Methylation: Molecular Biology and Biological Significance, (eds.) Jost, J.P., Saluz, H.P. (Birkhäuser Verlag Basel/Switzerland) pp. 145–168.

    Google Scholar 

  39. Borer, R.A., Lehner, C.F., Eppenberger, H.M., and Nigg, E.A. (1989).Cell 56:379–390.

    Article  PubMed  CAS  Google Scholar 

  40. Liu, Q.R., and Chan, P.K. (1993).DNA Cell Biol. 12:149–156.

    PubMed  CAS  Google Scholar 

  41. Bourbon, H.-M., and Amalric F. (1991):Gene 101:273–278.

    Article  CAS  Google Scholar 

  42. Cortese, J.F., Majewski, J.L., Crabb, D.W., Edenberg, H.J., and Yang, V.W. (1994).J. Biol. Chem. 269:21898–21906.

    PubMed  CAS  Google Scholar 

  43. Schmidt, M., and Migeon, B.R. (1990).Proc. Natl. Acad. Sci. U.S.A. 87:3685–3689.

    PubMed  CAS  Google Scholar 

  44. Sharman, G.B. (1971).Nature 230:231–232.

    Article  PubMed  CAS  Google Scholar 

  45. Barr, M.L., and Bertram, E.G. (1949).Nature 163:676–677.

    Google Scholar 

  46. Johnston, P.G., and Robinson, E.S. (1987).Chromosoma 95:419–423.

    Article  PubMed  CAS  Google Scholar 

  47. Yagil, G. (1991).Crit. Rev. Biochem. Mol. Biol. 26:475–559.

    PubMed  CAS  Google Scholar 

  48. Hardison, R., Chao, K.-M., Adamkiewicz, M., Price, D., Jackson, J., Zetgler, T., Stojanovic, N., and Miller, W. (1993).J. DNA Sequencing and Mapping 4:163–176.

    CAS  Google Scholar 

  49. Chen, A., Reyes, A., and Akeson, R. (1993).Gene 128:211–218.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, University of Technology, 2065, Sydney, New South Wales, Australia

    Suyinn Chong & Anita A. Piper

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  1. Suyinn Chong
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  2. Anita A. Piper
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The sequence data in this paper has been submitted to GenBank under the accession number U59897

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Chong, S., Piper, A.A. Methylation sensitive protein binding to an intragenic active X-specific methylated region in theM. robustus HPRT gene. Somat Cell Mol Genet 22, 461–476 (1996). https://doi.org/10.1007/BF02369438

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  • DOI: https://doi.org/10.1007/BF02369438

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