Advertisement

Somatic Cell and Molecular Genetics

, Volume 22, Issue 6, pp 435–441 | Cite as

Nuclease sensitivity of permeabilized cells confirms altered chromatin formation at the fragile X locus

  • Derek E. Eberhart
  • Stephen T. Warren
Article

Abstract

Fragile X syndrome is caused by the expansion and concomitant methylation of a CGG repeat in the 5′ untranslated region of the FMR1 gene which results in the transcriptional silencing of the FMR1 gene, delayed replication of the FMR1 locus, and the formation of a folate sensitive fragile site (FRAXA) at Xq27.3. The mechanism by which repeat expansion and methylation causes these changes is unknown. An in vivo system in which cells were permeabilized with lysophosphatidylcholine followed by digestion with MspI endonuclease was utilized to assess the chromatin conformation at the fragile X locus. The FMR1 gene was inaccessible to Mspl digestion in fragile X patients, but not in normal or carrier individuals, confirming that altered chromatin conformation results from the repeat expansion and methylation seen in fragile X syndrome.

Keywords

Folate Untranslated Region Permeabilized Cell Repeat Expansion Fragile Site 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Literature Cited

  1. 1.
    Warren, S.T., and Ashley, C.T. (1995).Annu. Rev. Neurosci. 18:77–99.CrossRefPubMedGoogle Scholar
  2. 2.
    Eberhart, D.E., and Warren, S.T.Cold Spring Harb. Symp. Quant. Biol. 61: in press.Google Scholar
  3. 3.
    Khandjian, E.W., Corbin, F., Woerly, S., and Rousseau, F. (1996).Nature Genet. 12:91–93.CrossRefPubMedGoogle Scholar
  4. 4.
    Tamamni, F., Meijer, N., Verheij, C., Willems, P.J., Galjaard, H., Oostra, B.A., and Hoogeveen, A.T. (1996).Hum. Mol. Genet. 5:809–813.Google Scholar
  5. 5.
    Eberhart, D.E., Malter, H.E., Feng, Y., and Warren, S.T. (1996).Hum. Mol. Genet. In press.Google Scholar
  6. 6.
    Ashley, C.T., Wilkmson, K.D., Remes, D., and Warren, S.T. (1993).Science 262:563–566.PubMedGoogle Scholar
  7. 7.
    Siomi, H., Siomi, M.C., Nussbaum, R.L., and Dreyfuss, G. (1993).Cell 74:291–298.CrossRefPubMedGoogle Scholar
  8. 8.
    Fu, Y.-H., Kuhl, D.P., Pizzuu, A., Pieretti, M., Sutcliffe, J.S., Richards, S., Verkerk, A.J.M.H., Holden, J.J.A., Fenwick, R.G., Jr., Warren, S.T., Oostra, B.A., Nelson, D.L., and Caskey, C.T. (1991).Cell 67:1047–1058.CrossRefPubMedGoogle Scholar
  9. 9.
    Oberlé, I., Rousseau, F., Heitz, D., Kretz, C., Devys, D., Hanauer, A., Boué, J., Bertheas, M.F., and Mandel, J.L. (1991).Science 252:1097–1102.PubMedGoogle Scholar
  10. 10.
    Verkerk, A.J.M.H., Piereth, M., Sutcliffe, J.S., Fu, Y.H., Kuhl, D.P.A., Pizutti, A., Remer, O., Richards, S., Victoria, M.F., Zhang, F., Eussen, B.E., van Ommen, G.J.B., Blonden, L.A.J., Riggins, G.J., Chastain, J.L., Kunst, C.B., Galjaard, H., Caskey, C.T., Nelson, D.L., Oostra, B.A., and Warren, S.T. (1991).Cell 65:905–914.CrossRefPubMedGoogle Scholar
  11. 11.
    Sutcliffe, J.S., Nelson, D.L., Zhang, F., Pieretti, M., Caskey, C.T., Saxe, D., and Warren, S.T. (1992).Hum. Mol. Genet. 1:397–400.PubMedGoogle Scholar
  12. 12.
    Hornstra, I.K., Nelson, D.L., Warren, S.T., and Yang, T.P. (1993).Hum. Mol. Genet. 2:1659–1665.PubMedGoogle Scholar
  13. 13.
    Ashley, C.T., Sutcliffe, J.S., Kunst, C.B., Leiner, H.A., Eichler, E.E., Nelson, D.L., and Warren, S.T. (1993).Nature Genet. 4:244–251.CrossRefPubMedGoogle Scholar
  14. 14.
    Snow, K., Doud, L.K., Hagerman, R., Pergolizzi, R.G., Erster, S.H., and Thibodeau, S.N. (1993).Am. J. Hum. Genet. 53:1217–1228.PubMedGoogle Scholar
  15. 15.
    Kunst, C.B., and Warren, S.T. (1994).Cell 77:853–861.CrossRefPubMedGoogle Scholar
  16. 16.
    Fisch, G.S., Snow, K., Thibodeau, S.N., Chalifaux, M., Holden, J.J.A., Nelson, D.L., Howard-Peebles, P.N., and Maddalena, A. (1995).Am. J. Hum. Genet. 56:1147–1155.PubMedGoogle Scholar
  17. 17.
    Hansen, R.S., Canfield, T.K., Lamb, M.M., Gartler, S.M., and Laird, C.D. (1993).Cell 73:1403–1409.CrossRefPubMedGoogle Scholar
  18. 18.
    Sutherland, G.R. (1977).Science 197:265–266.PubMedGoogle Scholar
  19. 19.
    Ashley, C.T., and Warren, S.T. (1995).Annu. Rev. Genet. 29:703–728.CrossRefPubMedGoogle Scholar
  20. 20.
    Gacy, A.M., Goellner, G., Juranie, N., Macura, S., and McMurray, C.T. (1995).Cell 81:533–540.PubMedGoogle Scholar
  21. 21.
    Fry, M., and Loeb, L.A. (1994).Proc. Natl. Acad. Sci. U.S.A. 91:4950–4954.PubMedGoogle Scholar
  22. 22.
    Luo, S., Robinson, J.C., Reiss, A.L., and Migeon, B.R. (1993):Somat. Cell Mol. Genet. 19:393–404.CrossRefPubMedGoogle Scholar
  23. 23.
    Pfeifer, G.P., and Riggs, A.D. (1991).Genes Dev. 5:1102–1113.PubMedGoogle Scholar
  24. 24.
    Taljanidisz, J., Popowski, J., and Sarkar, N. (1989).Mol. Cell. Biol. 9:2881–2889.PubMedGoogle Scholar
  25. 25.
    Pieretti, M., Zhang, F., Fu, Y.H., Warren, S.T., Oostra, B.A., Caskey, C.T., and Nelson, D.L. (1991).Cell 66:817–822.CrossRefPubMedGoogle Scholar
  26. 26.
    Fraser, N.J., Boyd, Y., and Craig, I. (1989).Genomics 5:144–148.CrossRefPubMedGoogle Scholar
  27. 27.
    Eberhart, D.E., and Warren, S.T. (1993).Am. J. Hum. Genet. 53 (suppl.):678 (abstract).Google Scholar
  28. 28.
    Ymer, S., and Jans, D.A. (1996).Biotechniques 20:834–840.PubMedGoogle Scholar
  29. 29.
    Warren, S.T., and Nelson, D.L. (1994).JAMA 271:536–542.CrossRefPubMedGoogle Scholar
  30. 30.
    Kirchgessner, C.U., Warren, S.T., and Willard, H.L., (1995).J. Med. Genet. 32:925–929.PubMedGoogle Scholar
  31. 31.
    Rhodes, K., Rippe, R.A., Umezawa, A., Nehls, M., Brenner, D.A., and Breindl, M. (1994).Mol. Cell. Biol. 14:5950–5960.PubMedGoogle Scholar
  32. 32.
    Pieper, R.O., Patel, S., Ting, S.A., Futscher, B.W., and Costello, J.F. (1996).J. Biol. Chem. 271:13916–13924.PubMedGoogle Scholar
  33. 33.
    Antequera, F., Boyes, J., and Bird, A. (1990).Cell 62:503–514.CrossRefPubMedGoogle Scholar
  34. 34.
    Keshet, L., Lieman-Hurwitz, J., and Cedar, H. (1986).Cell 44:535–543.CrossRefPubMedGoogle Scholar
  35. 35.
    Boyes, J., and Bird, A. (1991).Cell 64:1123–1134.CrossRefPubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  • Derek E. Eberhart
    • 1
  • Stephen T. Warren
    • 1
  1. 1.Howard Hughes Medical Institute and Departments of Biochemistry and PediatricsEmory University School of MedicineAtlanta

Personalised recommendations