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. 1978 Aug;5(8):2999–3012. doi: 10.1093/nar/5.8.2999

Transcription of nucleosomes from human chromatin.

P A Shaw, C G Sahasrabuddhe, H G Hodo 3rd, G F Saunders
PMCID: PMC342222  PMID: 693325

Abstract

Nucleosomes (chromatin subunits) prepared by micrococcal nuclease digestion of human nuclei are similar in histone content but substantially reduced in non-histone proteins as compared to undigested chromatin. Chromatin transcription experiments indicate that the DNA in the nucleosomes is accessible to DNA-dependent RNA polymerase in vitro. The template capacities of chromatin and nucleosomes are 1.5 and 10%, respectively, relative to high molecular weight DNA, with intermediate values for oligonucleosomes. Three distinct sizes of transcripts, 150, 120 and 95 nucleotides in length, are obtained when nucleosomes are used as templates. However, when nucleosomal DNA is used as a template, the predominant size of transcripts is 150 nucleotides. When oligonucleosomes are used as templates longer transcripts are obtained. This indicates that RNA polymerase can transcribe the DNA contained in the nucleosomes.

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Selected References

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  1. Bautz E. K., Dunn J. J. DNA-dependent RNA polymerase from phage T4 infected E. coli: an enzyme missing a factor required for transcription of T4 DNA. Biochem Biophys Res Commun. 1969 Jan 27;34(2):230–237. doi: 10.1016/0006-291x(69)90636-6. [DOI] [PubMed] [Google Scholar]
  2. Compton J. L., Bellard M., Chambon P. Biochemical evidence of variability in the DNA repeat length in the chromatin of higher eukaryotes. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4382–4386. doi: 10.1073/pnas.73.12.4382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. FLECK A., MUNRO H. N. The precision of ultraviolet absorption measurements in the Schmidt-Thannhauser procedure for nucleic acid estimation. Biochim Biophys Acta. 1962 May 14;55:571–583. doi: 10.1016/0006-3002(62)90836-3. [DOI] [PubMed] [Google Scholar]
  4. Foe V. E., Wilkinson L. E., Laird C. D. Comparative organization of active transcription units in Oncopeltus fasciatus. Cell. 1976 Sep;9(1):131–146. doi: 10.1016/0092-8674(76)90059-3. [DOI] [PubMed] [Google Scholar]
  5. Griffith J. D. Chromatin structure: deduced from a minichromosome. Science. 1975 Mar 28;187(4182):1202–1203. doi: 10.1126/science.187.4182.1202. [DOI] [PubMed] [Google Scholar]
  6. Hall M. R. Transcription and reisolation of the simian virus 40 nucleoprotein complex. Biochem Biophys Res Commun. 1977 Jun 6;76(3):698–704. doi: 10.1016/0006-291x(77)91556-x. [DOI] [PubMed] [Google Scholar]
  7. Hanlon S., Johnson R. S., Wolf B., Chan A. Mixed conformations of deoxyribonucleic acid in chromatin: a preliminary report. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3263–3267. doi: 10.1073/pnas.69.11.3263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kirby K. S., Cook E. A. Isolation of deoxyribonucleic acid from mammalian tissues. Biochem J. 1967 Jul;104(1):254–257. doi: 10.1042/bj1040254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kornberg R. D. Chromatin structure: a repeating unit of histones and DNA. Science. 1974 May 24;184(4139):868–871. doi: 10.1126/science.184.4139.868. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. 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]
  12. Langmore J. P., Wooley J. C. Chromatin architecture: investigation of a subunit of chromatin by dark field electron microscopy. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2691–2695. doi: 10.1073/pnas.72.7.2691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mandel R., Fasman G. D. Chromatin and nucleosome structure. Nucleic Acids Res. 1976 Aug;3(8):1839–1855. doi: 10.1093/nar/3.8.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Marushige K., Bonner J. Template properties of liver chromatin. J Mol Biol. 1966 Jan;15(1):160–174. doi: 10.1016/s0022-2836(66)80218-8. [DOI] [PubMed] [Google Scholar]
  16. Maurer H. R., Chalkley G. R. Some properties of a nuclear binding site of estradiol. J Mol Biol. 1967 Aug 14;27(3):431–441. doi: 10.1016/0022-2836(67)90049-6. [DOI] [PubMed] [Google Scholar]
  17. Olins A. L., Olins D. E. Spheroid chromatin units (v bodies). Science. 1974 Jan 25;183(4122):330–332. doi: 10.1126/science.183.4122.330. [DOI] [PubMed] [Google Scholar]
  18. Oosterhof D. K., Hozier J. C., Rill R. L. Nucleas action on chromatin: evidence for discrete, repeated nucleoprotein units along chromatin fibrils. Proc Natl Acad Sci U S A. 1975 Feb;72(2):633–637. doi: 10.1073/pnas.72.2.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Oudet P., Gross-Bellard M., Chambon P. Electron microscopic and biochemical evidence that chromatin structure is a repeating unit. Cell. 1975 Apr;4(4):281–300. doi: 10.1016/0092-8674(75)90149-x. [DOI] [PubMed] [Google Scholar]
  20. Sahasrabuddhe C. G., Saunders G. F. Salt-induced structural changes in nucleosomes. Nucleic Acids Res. 1977 Apr;4(4):853–866. doi: 10.1093/nar/4.4.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Sawada H., Crain W. R., Saunders G. F. Transcription of chromatin from two classes of human leukemic leukocytes. Biochim Biophys Acta. 1972 Nov 9;281(4):643–651. doi: 10.1016/0005-2787(72)90161-x. [DOI] [PubMed] [Google Scholar]
  23. Sawada H., Crain W. R., Saunders G. F. Transcription of chromatin from two classes of human leukemic leukocytes. Biochim Biophys Acta. 1972 Nov 9;281(4):643–651. doi: 10.1016/0005-2787(72)90161-x. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Tsai M. J., Saunders G. F. Transcription of chromatin by human RNA polymerase. Biochem Biophys Res Commun. 1973 Apr 2;51(3):756–765. doi: 10.1016/0006-291x(73)91380-6. [DOI] [PubMed] [Google Scholar]
  26. Weintraub H., Worcel A., Alberts B. A model for chromatin based upon two symmetrically paired half-nucleosomes. Cell. 1976 Nov;9(3):409–417. doi: 10.1016/0092-8674(76)90085-4. [DOI] [PubMed] [Google Scholar]
  27. van Bruggen E. F., Arnberg A. C., van Holde K. E., Sahasrabuddhe C. G., Shaw B. R. Electron microscopy of chromatin subunit particles. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1365–1370. doi: 10.1016/0006-291x(74)90348-9. [DOI] [PubMed] [Google Scholar]

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