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. 1976 Nov;73(11):3966–3970. doi: 10.1073/pnas.73.11.3966

Selective digestion of transcriptionally active ovalbumin genes from oviduct nuclei.

A Garel, R Axel
PMCID: PMC431285  PMID: 1069279

Abstract

Analysis of the DNA of isolated nucleosomes suggests that virtually all genomic DNA sequences are organized in this basic chromatin subunit. In this report, we demonstrate that although histones reside on the transcriptionally active ovalbumin genes in the oviduct, the organization of proteins about this gene renders it highly sensitive to deoxyribonuclease I (deoxyribonucleate 5'-oligonucleotidohydrolase, EC 3.1.4.5). Treatment of oviduct nuclei from the laying hen with pancreatic deoxyribonuclease I results in the preferential digestion of over 70% of the ovalbumin sequences when only 10% of the total nuclear DNA has been solubilized. Treatment of liver nuclei does not reveal selective sensitivity of these genes to DNase I. Furthermore, regions of DNA not actively transcribed, such as the endogenous leukosis virus genes in the oviduct, are not selectively degraded by this enzyme. Similar digestions with micrococcal nuclease, however, reveal no specific digestion of transcriptionally active chromatin. These data confirm the observations of H. Weintraub and M. Groudine [(1976) Science 193, 848-856] and suggest we are dealing with an aspect of structure that may be necessary to permit transcription of the chromatin complex.

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Selected References

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

  1. Axel R., Cedar H., Felsenfeld G. Synthesis of globin ribonucleic acid from duck-reticulocyte chromatin in vitro. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2029–2032. doi: 10.1073/pnas.70.7.2029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axel R., Cedar H., Felsenfield G. The structure of the globin genes in chromatin. Biochemistry. 1975 Jun 3;14(11):2489–2495. doi: 10.1021/bi00682a031. [DOI] [PubMed] [Google Scholar]
  3. Axel R. Cleavage of DNA in nuclei and chromatin with staphylococcal nuclease. Biochemistry. 1975 Jul;14(13):2921–2925. doi: 10.1021/bi00684a020. [DOI] [PubMed] [Google Scholar]
  4. Axel R., Melchior W., Jr, Sollner-Webb B., Felsenfeld G. Specific sites of interaction between histones and DNA in chromatin. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4101–4105. doi: 10.1073/pnas.71.10.4101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berkowitz E. M., Doty P. Chemical and physical properties of fractionated chromatin. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3328–3332. doi: 10.1073/pnas.72.9.3328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
  7. Camerini-Otero R. D., Sollner-Webb B., Felsenfeld G. The organization of histones and DNA in chromatin: evidence for an arginine-rich histone kernel. Cell. 1976 Jul;8(3):333–347. doi: 10.1016/0092-8674(76)90145-8. [DOI] [PubMed] [Google Scholar]
  8. Gilmour R. S., Paul J. Tissue-specific transcription of the globin gene in isolated chromatin. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3440–3442. doi: 10.1073/pnas.70.12.3440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gottesfeld J. M., Murphy R. F., Bonner J. Structure of transcriptionally active chromatin. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4404–4408. doi: 10.1073/pnas.72.11.4404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harris S. E., Schwartz R. J., Tsai M. J., O'Malley B. W., Roy A. K. Effect of estrogen on gene expression in the chick oviduct. In vitro transcription of the ovalbumin gene in chromatin. J Biol Chem. 1976 Jan 25;251(2):524–529. [PubMed] [Google Scholar]
  11. Hewish D. R., Burgoyne L. A. Chromatin sub-structure. The digestion of chromatin DNA at regularly spaced sites by a nuclear deoxyribonuclease. Biochem Biophys Res Commun. 1973 May 15;52(2):504–510. doi: 10.1016/0006-291x(73)90740-7. [DOI] [PubMed] [Google Scholar]
  12. Jacquet M., Groner Y., Monroy G., Hurwitz J. The in vitro synthesis of avian myeloblastosis viral RNA sequences. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3045–3049. doi: 10.1073/pnas.71.8.3045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kornberg R. D., Thomas J. O. Chromatin structure; oligomers of the histones. Science. 1974 May 24;184(4139):865–868. doi: 10.1126/science.184.4139.865. [DOI] [PubMed] [Google Scholar]
  14. Lacy E., Axel R. Analysis of DNA of isolated chromatin subunits. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3978–3982. doi: 10.1073/pnas.72.10.3978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maniatis T., Jeffrey A., van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. doi: 10.1021/bi00688a010. [DOI] [PubMed] [Google Scholar]
  16. Noll M. Internal structure of the chromatin subunit. Nucleic Acids Res. 1974 Nov;1(11):1573–1578. doi: 10.1093/nar/1.11.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Noll M. Subunit structure of chromatin. Nature. 1974 Sep 20;251(5472):249–251. doi: 10.1038/251249a0. [DOI] [PubMed] [Google Scholar]
  18. Olins A. L., Carlson R. D., Olins D. E. Visualization of chromatin substructure: upsilon bodies. J Cell Biol. 1975 Mar;64(3):528–537. doi: 10.1083/jcb.64.3.528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Palmiter R. D. Quantitation of parameters that determine the rate of ovalbumin synthesis. Cell. 1975 Mar;4(3):189–189. doi: 10.1016/0092-8674(75)90167-1. [DOI] [PubMed] [Google Scholar]
  20. Sahasrabuddhe C. G., Van Holde K. E. The effect of trypsin on nuclease-resistant chromatin fragments. J Biol Chem. 1974 Jan 10;249(1):152–156. [PubMed] [Google Scholar]
  21. Shoyab M., Baluda M. A., Evans R. Acquisition of new DNA sequences after infection of chicken cells with avian myeloblastosis virus. J Virol. 1974 Feb;13(2):331–339. doi: 10.1128/jvi.13.2.331-339.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Steggles A. W., Wilson G. N., Kantor J. A., Picciano D. J., Falvey A. K., Anderson W. F. Cell-free transcription of mammalian chromatin: transcription of globin messenger RNA sequences from bone-marrow chromatin with mammalian RNA polymerase. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1219–1223. doi: 10.1073/pnas.71.4.1219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tien Kuo M., Sahasrabuddhe C. G., Saunders G. F. Presence of messenger specifying sequences in the DNA of chromatin subunits. Proc Natl Acad Sci U S A. 1976 May;73(5):1572–1575. doi: 10.1073/pnas.73.5.1572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]

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