Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1983 Feb 11;11(3):753–772. doi: 10.1093/nar/11.3.753

The nuclease sensitivity of active genes.

R H Nicolas, C A Wright, P N Cockerill, J A Wyke, G H Goodwin
PMCID: PMC325751  PMID: 6300766

Abstract

Brief micrococcal nuclease digestion of chick embryonic red blood cells results in preferential excision and solubilization of monomer nucleosomes associated with beta-globin sequences and also 5'-sequences flanking the beta-globin gene. Both regions are DNAse-I sensitive in nuclei. Such salt-soluble nucleosomes are enriched in all four major HMG proteins but HMG1 and 2 are only weakly associated. These nucleosomes appear to have lost much of the DNAse-I sensitivity of active genes. The HMG14 and 17-containing salt-soluble nucleosomes separated by electrophoresis are not DNAse-I sensitive and contain inactive gene sequences as well as active sequences. Reconstitution of HMG proteins onto bulk nucleosomes or chromatin failed to reveal an HMG-dependent sensitivity of active genes as assayed by dot-blot hybridization and it was found that the DNAse-I sensitivity of ASV proviral sequences as assayed by dot-blot hybridization was not HMG-dependent. These results indicate that higher order chromatin structures might be responsible for nuclease sensitivity of active genes.

Full text

PDF
753

Images in this article

Selected References

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

  1. Albanese I., Weintraub H. Electrophoretic separation of a class of nucleosomes enriched in HMG 14 and 17 and actively transcribed globin genes. Nucleic Acids Res. 1980 Jun 25;8(12):2787–2805. doi: 10.1093/nar/8.12.2787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albright S. C., Wiseman J. M., Lange R. A., Garrard W. T. Subunit structures of different electrophoretic forms of nucleosomes. J Biol Chem. 1980 Apr 25;255(8):3673–3684. [PubMed] [Google Scholar]
  3. Barsoum J., Levinger L., Varshavsky A. On the chromatin structure of the amplified, transcriptionally active gene for dihydrofolate reductase in mouse cells. J Biol Chem. 1982 May 10;257(9):5274–5282. [PubMed] [Google Scholar]
  4. Bellard M., Gannon F., Chambon P. Nucleosome structure III: the structure and transcriptional activity of the chromatin containing the ovalbumin and globin genes in chick oviduct nuclei. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):779–791. doi: 10.1101/sqb.1978.042.01.078. [DOI] [PubMed] [Google Scholar]
  5. Bloom K. S., Anderson J. N. Conformation of ovalbumin and globin genes in chromatin during differential gene expression. J Biol Chem. 1979 Oct 25;254(20):10532–10539. [PubMed] [Google Scholar]
  6. Bloom K. S., Anderson J. N. Fractionation of hen oviduct chromatin into transcriptionally active and inactive regions after selective micrococcal nuclease digestion. Cell. 1978 Sep;15(1):141–150. doi: 10.1016/0092-8674(78)90090-9. [DOI] [PubMed] [Google Scholar]
  7. Chiswell D. J., Gillespie D. A., Wyke J. A. The changes in proviral chromatin that accompany morphological variation in avian sarcoma virus-infected rat cells. Nucleic Acids Res. 1982 Jul 10;10(13):3967–3980. doi: 10.1093/nar/10.13.3967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DeLorbe W. J., Luciw P. A., Goodman H. M., Varmus H. E., Bishop J. M. Molecular cloning and characterization of avian sarcoma virus circular DNA molecules. J Virol. 1980 Oct;36(1):50–61. doi: 10.1128/jvi.36.1.50-61.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dolan M., Sugarman B. J., Dodgson J. B., Engel J. D. Chromosomal arrangement of the chicken beta-type globin genes. Cell. 1981 Jun;24(3):669–677. doi: 10.1016/0092-8674(81)90093-3. [DOI] [PubMed] [Google Scholar]
  10. Egan P. A., Levy-Wilson B. Structure of transcriptionally active and inactive nucleosomes from butyrate-treated and control HeLa cells. Biochemistry. 1981 Jun 23;20(13):3695–3702. doi: 10.1021/bi00516a005. [DOI] [PubMed] [Google Scholar]
  11. Gabrielli F., Hancock R., Faber A. J. Characterisation of a chromatin fraction bearing pulse-labelled RNA. 2. Quantification of histones and high-mobility-group proteins. Eur J Biochem. 1981 Nov;120(2):363–369. doi: 10.1111/j.1432-1033.1981.tb05713.x. [DOI] [PubMed] [Google Scholar]
  12. Goldsmith M. E. Release of a globin gene enriched chromatin fraction from chicken erythrocyte nuclei following DNase II digestion. Nucleic Acids Res. 1981 Dec 11;9(23):6471–6485. doi: 10.1093/nar/9.23.6471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goodwin G. H., Mathew C. G., Wright C. A., Venkov C. D., Johns E. W. Analysis of the high mobility group proteins associated with salt-soluble nucleosomes. Nucleic Acids Res. 1979 Dec 11;7(7):1815–1835. doi: 10.1093/nar/7.7.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goodwin G. H., Wright C. A., Johns E. W. The characterisation of 1SF monomer nucleosomes from hen oviduct and the partial characterisation of a third HMG14/17-like in such nucleosomes. Nucleic Acids Res. 1981 Jun 25;9(12):2761–2775. doi: 10.1093/nar/9.12.2761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Humphries P., Cochet M., Krust A., Gerlinger P., Kourilsky P., Chambon P. Molecular cloning of extensive sequences of the in vitro synthesized chicken ovalbumin structural gene. Nucleic Acids Res. 1977 Jul;4(7):2389–2406. doi: 10.1093/nar/4.7.2389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Larsen A., Weintraub H. An altered DNA conformation detected by S1 nuclease occurs at specific regions in active chick globin chromatin. Cell. 1982 Jun;29(2):609–622. doi: 10.1016/0092-8674(82)90177-5. [DOI] [PubMed] [Google Scholar]
  17. Lawson G. M., Knoll B. J., March C. J., Woo S. L., Tsai M. J., O'Malley B. W. Definition of 5' and 3' structural boundaries of the chromatin domain containing the ovalbumin multigene family. J Biol Chem. 1982 Feb 10;257(3):1501–1507. [PubMed] [Google Scholar]
  18. Levinger L., Barsoum J., Varshavsky A. Two-dimensional hybridization mapping of nucleosomes. comparison of DNA and protein patterns. J Mol Biol. 1981 Mar 5;146(3):287–304. doi: 10.1016/0022-2836(81)90389-2. [DOI] [PubMed] [Google Scholar]
  19. Levy B. W., Connor W., Dixon G. H. A subset of trout testis nucleosomes enriched in transcribed DNA sequences contains high mobility group proteins as major structural components. J Biol Chem. 1979 Feb 10;254(3):609–620. [PubMed] [Google Scholar]
  20. Mardian J. K., Paton A. E., Bunick G. J., Olins D. E. Nucleosome cores have two specific binding sites for nonhistone chromosomal proteins HMG 14 and HMG 17. Science. 1980 Sep 26;209(4464):1534–1536. doi: 10.1126/science.7433974. [DOI] [PubMed] [Google Scholar]
  21. Mathew C. G., Goodwin G. H., Igo-Kemenes T., Johns E. W. The protein composition of rat satellite chromatin. FEBS Lett. 1981 Mar 9;125(1):25–29. doi: 10.1016/0014-5793(81)80988-x. [DOI] [PubMed] [Google Scholar]
  22. Mathew C. G., Goodwin G. H., Wright C. A., Johns E. W. The high mobility group proteins and transcribed nucleosomes. Cell Biol Int Rep. 1981 Jan;5(1):37–43. doi: 10.1016/0309-1651(81)90155-7. [DOI] [PubMed] [Google Scholar]
  23. McGhee J. D., Wood W. I., Dolan M., Engel J. D., Felsenfeld G. A 200 base pair region at the 5' end of the chicken adult beta-globin gene is accessible to nuclease digestion. Cell. 1981 Nov;27(1 Pt 2):45–55. doi: 10.1016/0092-8674(81)90359-7. [DOI] [PubMed] [Google Scholar]
  24. Parker M., Needham M., White R. Prostatic steroid binding protein: gene duplication and steroid binding. Nature. 1982 Jul 1;298(5869):92–94. doi: 10.1038/298092a0. [DOI] [PubMed] [Google Scholar]
  25. Reudelhuber T. L., Ball D. J., Davis A. H., Garrard W. T. Transferring DNA from electrophoretically resolved nucleosomes to diazobenzyloxymethyl cellulose: properties of nucleosomes along mouse satellite DNA. Nucleic Acids Res. 1982 Feb 25;10(4):1311–1325. doi: 10.1093/nar/10.4.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sandeen G., Wood W. I., Felsenfeld G. The interaction of high mobility proteins HMG14 and 17 with nucleosomes. Nucleic Acids Res. 1980 Sep 11;8(17):3757–3778. doi: 10.1093/nar/8.17.3757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Seed B. Diazotizable arylamine cellulose papers for the coupling and hybridization of nucleic acids. Nucleic Acids Res. 1982 Mar 11;10(5):1799–1810. doi: 10.1093/nar/10.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Senear A. W., Palmiter R. D. Multiple structural features are responsible for the nuclease sensitivity of the active ovalbumin gene. J Biol Chem. 1981 Feb 10;256(3):1191–1198. [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. 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. 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]
  32. Todd R. D., Garrard W. T. Two-dimensional electrophoretic analysis of polynucleosomes. J Biol Chem. 1977 Jul 10;252(13):4729–4738. [PubMed] [Google Scholar]
  33. Villeponteau B., Landes G. M., Pankratz M. J., Martinson H. G. The chicken beta globin gene region. Delineation of transcription units and developmental regulation of interspersed DNA repeats. J Biol Chem. 1982 Sep 25;257(18):11015–11023. [PubMed] [Google Scholar]
  34. 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]
  35. Weintraub H., Groudine M. Chromosomal subunits in active genes have an altered conformation. Science. 1976 Sep 3;193(4256):848–856. doi: 10.1126/science.948749. [DOI] [PubMed] [Google Scholar]
  36. Weintraub H., Larsen A., Groudine M. Alpha-Globin-gene switching during the development of chicken embryos: expression and chromosome structure. Cell. 1981 May;24(2):333–344. doi: 10.1016/0092-8674(81)90323-8. [DOI] [PubMed] [Google Scholar]
  37. Weisbrod S. Active chromatin. Nature. 1982 May 27;297(5864):289–295. doi: 10.1038/297289a0. [DOI] [PubMed] [Google Scholar]
  38. Weisbrod S., Groudine M., Weintraub H. Interaction of HMG 14 and 17 with actively transcribed genes. Cell. 1980 Jan;19(1):289–301. doi: 10.1016/0092-8674(80)90410-9. [DOI] [PubMed] [Google Scholar]
  39. Weisbrod S., Weintraub H. Isolation of actively transcribed nucleosomes using immobilized HMG 14 and 17 and an analysis of alpha-globin chromatin. Cell. 1981 Feb;23(2):391–400. doi: 10.1016/0092-8674(81)90134-3. [DOI] [PubMed] [Google Scholar]
  40. Wood W. I., Felsenfeld G. Chromatin structure of the chicken beta-globin gene region. Sensitivity to DNase I, micrococcal nuclease, and DNase II. J Biol Chem. 1982 Jul 10;257(13):7730–7736. [PubMed] [Google Scholar]
  41. Wood W. I., Nickol J., Felsenfeld G. Repeated sequence organization and RNA transcription map of the chicken adult beta-globin gene region. J Biol Chem. 1981 Feb 25;256(4):1502–1506. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES