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. 1981 Apr 24;9(8):1841–1851. doi: 10.1093/nar/9.8.1841

Modification of the methylation pattern in the vicinity of the chicken globin genes in avian erythroblastosis virus transformed cells.

L Marcaud, C A Reynaud, A Therwath, K Scherrer
PMCID: PMC326807  PMID: 6264396

Abstract

Having previously found a reduced transcription of globin genes and an abortive processing of the already transcribed globin pre-mRNA in Avian Erythroblastosis Virus (AEV) transformed cells (1), we compared the genomic DNA of these cells with that of normal chicken erythroblasts, using 32-P-labelled cDNA probes specific for the beta, alpha A and alpha D globin sequences. Restriction endonuclease digestion, electrophoresis of digests in agarose gels, Southern blotting and hybridization were carried out. Our results show that the overall genome organization is not disturbed in the immediate neighbourhood of the adult globin genes; the observed restriction fragments are identical for both DNAs after EcoRI, HindIII, BamHI and XbaI digestion, using the beta, alpha A and alpha D globin cDNA probes. However, we observe specific modifications at some methylation sites in the beta, beta-like and alpha D regions: after HpaII or MspI digestion in the alpha D region and after HhaI digestion in the beta and beta-like region, heavier bands appear in the transformed cell DNA in addition to the ones observed in normal DNA. This implies that, at some specific sites, the transformed cell DNA is more methylated than the normal erythroblast DNA. The possible significance of this observation is discussed.

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Selected References

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

  1. Beug H., von Kirchbach A., Döderlein G., Conscience J. F., Graf T. Chicken hematopoietic cells transformed by seven strains of defective avian leukemia viruses display three distinct phenotypes of differentiation. Cell. 1979 Oct;18(2):375–390. doi: 10.1016/0092-8674(79)90057-6. [DOI] [PubMed] [Google Scholar]
  2. Buss L. W. Competitive intransitivity and size-frequency distributions of interacting populations. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5355–5359. doi: 10.1073/pnas.77.9.5355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chan L. N. Transport of globin mRNA from nucleus into cytoplasm in differentiating embryonic red blood cells. Nature. 1976 May 13;261(5556):157–159. doi: 10.1038/261157a0. [DOI] [PubMed] [Google Scholar]
  4. Cohen J. C. Methylation of milk-borne and genetically transmitted mouse mammary tumor virus proviral DNA. Cell. 1980 Mar;19(3):653–662. doi: 10.1016/s0092-8674(80)80042-0. [DOI] [PubMed] [Google Scholar]
  5. Cummings I. W., Liu A. Y., Salser W. A. Identification of a new chicken alpha-globin structural gene by complementary DNA cloning. Nature. 1978 Nov 23;276(5686):418–419. doi: 10.1038/276418a0. [DOI] [PubMed] [Google Scholar]
  6. Deacon N. J., Shine J., Naora H. Complete nucleotide sequence of a cloned chicken alpha-globin cDNA. Nucleic Acids Res. 1980 Mar 25;8(6):1187–1199. doi: 10.1093/nar/8.6.1187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dodgson J. B., Strommer J., Engel J. D. Isolation of the chicken beta-globin gene and a linked embryonic beta-like globin gene from a chicken DNA recombinant library. Cell. 1979 Aug;17(4):879–887. doi: 10.1016/0092-8674(79)90328-3. [DOI] [PubMed] [Google Scholar]
  8. Engel J. D., Dodgson J. B. Analysis of the adult and embryonic chicken globin genes in chromosomal DNA. J Biol Chem. 1978 Nov 25;253(22):8239–8246. [PubMed] [Google Scholar]
  9. Engel J. D., Dodgson J. B. Analysis of the closely linked adult chicken alpha-globin genes in recombinant DNAs. Proc Natl Acad Sci U S A. 1980 May;77(5):2596–2600. doi: 10.1073/pnas.77.5.2596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gazzolo L., Samarut J., Bouabdelli M., Blanchet J. P. Early precursors in the erythroid lineage are the specific target cells of avian erythroblastosis virus in vitro. Cell. 1980 Dec;22(3):683–691. doi: 10.1016/0092-8674(80)90544-9. [DOI] [PubMed] [Google Scholar]
  11. Ginder G. D., Wood W. I., Felsenfeld G. Isolation and characterization of recombinant clones containing the chicken adult beta-globin gene. J Biol Chem. 1979 Sep 10;254(17):8099–8102. [PubMed] [Google Scholar]
  12. Groudine M., Weintraub H. Rous sarcoma virus activates embryonic globin genes in chicken fibroblasts. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4464–4468. doi: 10.1073/pnas.72.11.4464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Guntaka R. V., Rao P. Y., Mitsialis S. A., Katz R. Modification of avian sarcoma proviral DNA sequences in nonpermissive XC cells but not in permissive chicken cells. J Virol. 1980 May;34(2):569–572. doi: 10.1128/jvi.34.2.569-572.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hughes S. H., Payvar F., Spector D., Schimke R. T., Robinson H. L., Payne G. S., Bishop J. M., Varmus H. E. Heterogeneity of genetic loci in chickens: analysis of endogenous viral and nonviral genes by cleavage of DNA with restriction endonucleases. Cell. 1979 Oct;18(2):347–359. doi: 10.1016/0092-8674(79)90054-0. [DOI] [PubMed] [Google Scholar]
  15. Jones P. A., Taylor S. M. Cellular differentiation, cytidine analogs and DNA methylation. Cell. 1980 May;20(1):85–93. doi: 10.1016/0092-8674(80)90237-8. [DOI] [PubMed] [Google Scholar]
  16. Keane R. W., Lindblad P. C., Pierik L. T., Ingram V. M. Isolation and transformation of primary mesenchymal cells of the chick embryo. Cell. 1979 Aug;17(4):801–811. doi: 10.1016/0092-8674(79)90320-9. [DOI] [PubMed] [Google Scholar]
  17. Lai E. C., Woo S. L., Dugaiczyk A., O'Malley B. W. The ovalbumin gene: alleles created by mutations in the intervening sequences of the natural gene. Cell. 1979 Jan;16(1):201–211. doi: 10.1016/0092-8674(79)90201-0. [DOI] [PubMed] [Google Scholar]
  18. Mandel J. L., Chambon P. DNA methylation: organ specific variations in the methylation pattern within and around ovalbumin and other chicken genes. Nucleic Acids Res. 1979 Dec 20;7(8):2081–2103. doi: 10.1093/nar/7.8.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  20. McGhee J. D., Ginder G. D. Specific DNA methylation sites in the vicinity of the chicken beta-globin genes. Nature. 1979 Aug 2;280(5721):419–420. doi: 10.1038/280419a0. [DOI] [PubMed] [Google Scholar]
  21. Meunier-Rotival M., Cortadas J., Macaya G., Bernardi G. Isolation and organization of calf ribosomal DNA. Nucleic Acids Res. 1979;6(6):2109–2123. doi: 10.1093/nar/6.6.2109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Razin A., Riggs A. D. DNA methylation and gene function. Science. 1980 Nov 7;210(4470):604–610. doi: 10.1126/science.6254144. [DOI] [PubMed] [Google Scholar]
  23. Reynaud C. A., Ben Tahar S., Krust A., Amaral de Lima Franco M. P., Goldenberg S., Gannon F., Scherrer K. Restriction mapping of cDNA recombinants including the adult chicken and duck globin messenger sequences: a comparative study. Gene. 1980 Nov;11(3-4):259–269. doi: 10.1016/0378-1119(80)90066-9. [DOI] [PubMed] [Google Scholar]
  24. Reynaud C. A., Imaizumi-Scherrer M. T., Scherrer K. The size of the transcriptional units of the avian globin genes defined at the pre-messenger RNA level. J Mol Biol. 1980 Jul 15;140(4):481–504. doi: 10.1016/0022-2836(80)90267-3. [DOI] [PubMed] [Google Scholar]
  25. Richards R. I., Wells J. R. Chicken globin genes. Nucleotide sequence of cDNA clones coding for the alpha-globin expressed during hemolytic anemia. J Biol Chem. 1980 Oct 10;255(19):9306–9311. [PubMed] [Google Scholar]
  26. Rowley J. D. Letter: A new consistent chromosomal abnormality in chronic myelogenous leukaemia identified by quinacrine fluorescence and Giemsa staining. Nature. 1973 Jun 1;243(5405):290–293. doi: 10.1038/243290a0. [DOI] [PubMed] [Google Scholar]
  27. Shen C. K., Maniatis T. Tissue-specific DNA methylation in a cluster of rabbit beta-like globin genes. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6634–6638. doi: 10.1073/pnas.77.11.6634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Stalder J., Groudine M., Dodgson J. B., Engel J. D., Weintraub H. Hb switching in chickens. Cell. 1980 Apr;19(4):973–980. doi: 10.1016/0092-8674(80)90088-4. [DOI] [PubMed] [Google Scholar]
  30. Stalder J., Larsen A., Engel J. D., Dolan M., Groudine M., Weintraub H. Tissue-specific DNA cleavages in the globin chromatin domain introduced by DNAase I. Cell. 1980 Jun;20(2):451–460. doi: 10.1016/0092-8674(80)90631-5. [DOI] [PubMed] [Google Scholar]
  31. Therwath A., Scherrer K. Post-transcriptional suppression of globin gene expression in cells transformed by avian erythroblastosis virus. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3776–3780. doi: 10.1073/pnas.75.8.3776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vardimon L., Neumann R., Kuhlmann I., Sutter D., Doerfler W. DNA methylation and viral gene expression in adenovirus-transformed and -infected cells. Nucleic Acids Res. 1980 Jun 11;8(11):2461–2473. doi: 10.1093/nar/8.11.2461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]

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