Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Nov;81(21):6618–6622. doi: 10.1073/pnas.81.21.6618

DNA methylation and regulation of the human beta-globin-like genes in mouse erythroleukemia cells containing human chromosome 11.

T J Ley, Y L Chiang, D Haidaris, N P Anagnou, V L Wilson, W F Anderson
PMCID: PMC391981  PMID: 6208553

Abstract

The human beta-globin gene is expressed--but the human fetal (gamma) and embryonic (epsilon) globin genes are not--in an induced mouse erythroleukemia cell line (M11-X) that contains most of human chromosome 11. A 24-hr exposure of M11-X cells to 5-azacytidine before induction causes "global" DNA hypomethylation but selective activation of the human gamma-globin genes. Genomic DNA is remethylated 2-3 days after exposure to 5-azacytidine, but sequences near the human and mouse globin genes remain hypomethylated, suggesting that the remethylation process is inhibited in these regions.

Full text

PDF
6618

Images in this article

6619Image
on p.6619
6619Image
on p.6619
6619Image
on p.6619
6619Image
on p.6619
6621Image
on p.6621
6621Image
on p.6621
6621Image
on p.6621
6621Image
on p.6621

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Balcarek J. M., McMorris F. A. DNase I hypersensitive sites of globin genes of uninduced Friend erythroleukemia cells and changes during induction with dimethyl sulfoxide. J Biol Chem. 1983 Sep 10;258(17):10622–10628. [PubMed] [Google Scholar]
  2. Bell G. I., Pictet R. L., Rutter W. J., Cordell B., Tischer E., Goodman H. M. Sequence of the human insulin gene. Nature. 1980 Mar 6;284(5751):26–32. doi: 10.1038/284026a0. [DOI] [PubMed] [Google Scholar]
  3. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  4. Bestor T. H., Ingram V. M. Two DNA methyltransferases from murine erythroleukemia cells: purification, sequence specificity, and mode of interaction with DNA. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5559–5563. doi: 10.1073/pnas.80.18.5559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Busslinger M., Hurst J., Flavell R. A. DNA methylation and the regulation of globin gene expression. Cell. 1983 Aug;34(1):197–206. doi: 10.1016/0092-8674(83)90150-2. [DOI] [PubMed] [Google Scholar]
  6. Chao M. V., Mellon P., Charnay P., Maniatis T., Axel R. The regulated expression of beta-globin genes introduced into mouse erythroleukemia cells. Cell. 1983 Feb;32(2):483–493. doi: 10.1016/0092-8674(83)90468-3. [DOI] [PubMed] [Google Scholar]
  7. Charache S., Dover G., Smith K., Talbot C. C., Jr, Moyer M., Boyer S. Treatment of sickle cell anemia with 5-azacytidine results in increased fetal hemoglobin production and is associated with nonrandom hypomethylation of DNA around the gamma-delta-beta-globin gene complex. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4842–4846. doi: 10.1073/pnas.80.15.4842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chiang Y. L., Ley T. J., Sanders-Haigh L., Anderson W. F. Human globin gene expression in hybrid 2S MEL X human fibroblast cells. Somat Cell Mol Genet. 1984 Jul;10(4):399–407. doi: 10.1007/BF01535635. [DOI] [PubMed] [Google Scholar]
  9. Creusot F., Acs G., Christman J. K. Inhibition of DNA methyltransferase and induction of Friend erythroleukemia cell differentiation by 5-azacytidine and 5-aza-2'-deoxycytidine. J Biol Chem. 1982 Feb 25;257(4):2041–2048. [PubMed] [Google Scholar]
  10. DeSimone J., Heller P., Hall L., Zwiers D. 5-Azacytidine stimulates fetal hemoglobin synthesis in anemic baboons. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4428–4431. doi: 10.1073/pnas.79.14.4428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Deisseroth A., Hendrick D. Human alpha-globin gene expression following chromosomal dependent gene transfer into mouse erythroleukemia cells. Cell. 1978 Sep;15(1):55–63. doi: 10.1016/0092-8674(78)90082-x. [DOI] [PubMed] [Google Scholar]
  12. Felsenfeld G., McGhee J. Methylation and gene control. Nature. 1982 Apr 15;296(5858):602–603. doi: 10.1038/296602a0. [DOI] [PubMed] [Google Scholar]
  13. Ginder G. D., Whitters M. J., Pohlman J. K. Activation of a chicken embryonic globin gene in adult erythroid cells by 5-azacytidine and sodium butyrate. Proc Natl Acad Sci U S A. 1984 Jul;81(13):3954–3958. doi: 10.1073/pnas.81.13.3954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Groudine M., Kohwi-Shigematsu T., Gelinas R., Stamatoyannopoulos G., Papayannopoulou T. Human fetal to adult hemoglobin switching: changes in chromatin structure of the beta-globin gene locus. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7551–7555. doi: 10.1073/pnas.80.24.7551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Humphries R. K., Ley T., Turner P., Moulton A. D., Nienhuis A. W. Differences in human alpha-, beta- and delta-globin gene expression in monkey kidney cells. Cell. 1982 Aug;30(1):173–183. doi: 10.1016/0092-8674(82)90023-x. [DOI] [PubMed] [Google Scholar]
  16. Konkel D. A., Tilghman S. M., Leder P. The sequence of the chromosomal mouse beta-globin major gene: homologies in capping, splicing and poly(A) sites. Cell. 1978 Dec;15(4):1125–1132. doi: 10.1016/0092-8674(78)90040-5. [DOI] [PubMed] [Google Scholar]
  17. Lachman H. M., Mears J. G. DNase I hypersensitivity in the gamma globin gene locus of K562 cells. Nucleic Acids Res. 1983 Sep 10;11(17):6065–6077. doi: 10.1093/nar/11.17.6065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Letvin N. L., Linch D. C., Beardsley G. P., McIntyre K. W., Nathan D. G. Augmentation of fetal-hemoglobin production in anemic monkeys by hydroxyurea. N Engl J Med. 1984 Apr 5;310(14):869–873. doi: 10.1056/NEJM198404053101401. [DOI] [PubMed] [Google Scholar]
  19. Ley T. J., Anagnou N. P., Pepe G., Nienhuis A. W. RNA processing errors in patients with beta-thalassemia. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4775–4779. doi: 10.1073/pnas.79.15.4775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ley T. J., DeSimone J., Anagnou N. P., Keller G. H., Humphries R. K., Turner P. H., Young N. S., Keller P., Nienhuis A. W. 5-azacytidine selectively increases gamma-globin synthesis in a patient with beta+ thalassemia. N Engl J Med. 1982 Dec 9;307(24):1469–1475. doi: 10.1056/NEJM198212093072401. [DOI] [PubMed] [Google Scholar]
  21. Ley T. J., DeSimone J., Noguchi C. T., Turner P. H., Schechter A. N., Heller P., Nienhuis A. W. 5-Azacytidine increases gamma-globin synthesis and reduces the proportion of dense cells in patients with sickle cell anemia. Blood. 1983 Aug;62(2):370–380. [PubMed] [Google Scholar]
  22. Marks P. A., Rifkind R. A. Erythroleukemic differentiation. Annu Rev Biochem. 1978;47:419–448. doi: 10.1146/annurev.bi.47.070178.002223. [DOI] [PubMed] [Google Scholar]
  23. Mavilio F., Giampaolo A., Carè A., Migliaccio G., Calandrini M., Russo G., Pagliardi G. L., Mastroberardino G., Marinucci M., Peschle C. Molecular mechanisms of human hemoglobin switching: selective undermethylation and expression of globin genes in embryonic, fetal, and adult erythroblasts. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6907–6911. doi: 10.1073/pnas.80.22.6907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nudel U., Salmon J., Fibach E., Terada M., Rifkind R., Marks P. A., Bank A. Accumulation of alpha- and beta-globin messenger RNAs in mouse erythroleukemia cells. Cell. 1977 Oct;12(2):463–469. doi: 10.1016/0092-8674(77)90122-2. [DOI] [PubMed] [Google Scholar]
  25. Owerbach D., Bell G. I., Rutter W. J., Brown J. A., Shows T. B. The insulin gene is located on the short arm of chromosome 11 in humans. Diabetes. 1981 Mar;30(3):267–270. doi: 10.2337/diab.30.3.267. [DOI] [PubMed] [Google Scholar]
  26. Papayannopoulou T., Torrealba de Ron A., Veith R., Knitter G., Stamatoyannopoulos G. Arabinosylcytosine induces fetal hemoglobin in baboons by perturbing erythroid cell differentiation kinetics. Science. 1984 May 11;224(4649):617–619. doi: 10.1126/science.6200940. [DOI] [PubMed] [Google Scholar]
  27. Scott A. F., Phillips J. A., 3rd, Migeon B. R. DNA restriction endonuclease analysis for localization of human beta- and delta-globin genes on chromosome 11. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4563–4565. doi: 10.1073/pnas.76.9.4563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sheffery M., Rifkind R. A., Marks P. A. Murine erythroleukemia cell differentiation: DNase I hypersensitivity and DNA methylation near the globin genes. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1180–1184. doi: 10.1073/pnas.79.4.1180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  30. Tanaka K., Appella E., Jay G. Developmental activation of the H-2K gene is correlated with an increase in DNA methylation. Cell. 1983 Dec;35(2 Pt 1):457–465. doi: 10.1016/0092-8674(83)90179-4. [DOI] [PubMed] [Google Scholar]
  31. Taylor S. M., Jones P. A. Mechanism of action of eukaryotic DNA methyltransferase. Use of 5-azacytosine-containing DNA. J Mol Biol. 1982 Dec 15;162(3):679–692. doi: 10.1016/0022-2836(82)90395-3. [DOI] [PubMed] [Google Scholar]
  32. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tonkonow B. L., Hoffman R., Burger D., Elder J. T., Mazur E. M., Murnane M. J., Benz E. J., Jr Differing responses of globin and glycophorin gene expression to hemin in the human leukemia cell line K562. Blood. 1982 Apr;59(4):738–746. [PubMed] [Google Scholar]
  34. Torrealba-de Ron A. T., Papayannopoulou T., Knapp M. S., Fu M. F., Knitter G., Stamatoyannopoulos G. Perturbations in the erythroid marrow progenitor cell pools may play a role in the augmentation of HbF by 5-azacytidine. Blood. 1984 Jan;63(1):201–210. [PubMed] [Google Scholar]
  35. Waalwijk C., Flavell R. A. DNA methylation at a CCGG sequence in the large intron of the rabbit beta-globin gene: tissue-specific variations. Nucleic Acids Res. 1978 Dec;5(12):4631–4634. doi: 10.1093/nar/5.12.4631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weisbrod S. Active chromatin. Nature. 1982 May 27;297(5864):289–295. doi: 10.1038/297289a0. [DOI] [PubMed] [Google Scholar]
  37. Willing M. C., Nienhuis A. W., Anderson W. F. Selective activation of human beta-but not gamma-globin gene in human fibroblast x mouse erythroleukaemia cell hybrids. Nature. 1979 Feb 15;277(5697):534–538. doi: 10.1038/277534a0. [DOI] [PubMed] [Google Scholar]
  38. Wilson V. L., Jones P. A. Inhibition of DNA methylation by chemical carcinogens in vitro. Cell. 1983 Jan;32(1):239–246. doi: 10.1016/0092-8674(83)90514-7. [DOI] [PubMed] [Google Scholar]
  39. Wright S., deBoer E., Grosveld F. G., Flavell R. A. Regulated expression of the human beta-globin gene family in murine erythroleukaemia cells. Nature. 1983 Sep 22;305(5932):333–336. doi: 10.1038/305333a0. [DOI] [PubMed] [Google Scholar]
  40. de Martinville B., Giacalone J., Shih C., Weinberg R. A., Francke U. Oncogene from human EJ bladder carcinoma is located on the short arm of chromosome 11. Science. 1983 Feb 4;219(4584):498–501. doi: 10.1126/science.6297001. [DOI] [PubMed] [Google Scholar]
  41. van der Ploeg L. H., Flavell R. A. DNA methylation in the human gamma delta beta-globin locus in erythroid and nonerythroid tissues. Cell. 1980 Apr;19(4):947–958. doi: 10.1016/0092-8674(80)90086-0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES