Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1976 Sep;3(9):2315–2329. doi: 10.1093/nar/3.9.2315

Some properties of tobacco protoplast chromatin.

C Gigot, G Philipps, A Nicolaieff, L Hirth
PMCID: PMC343087  PMID: 967696

Abstract

Chromatin was prepared from tobacco-leaf protoplasts. Its solubility in increasing molarities of NaCl was studied and the structure of the soluble fraction observed by electron microscopy. We demonstrate that in plants, the DNA and histones are associated in beaded structures similar to those called omicron-bodies or nucleosomes in animal chromatin. The nucleosomes were associated with DNA in either compact or extended forms. The compact arrangement was predominant in the fraction solubilized between 0.1 and 0.4 M NaCl. The extended form, present at 0.5 and 0.6 M NaCl. showed DNA filaments of various lengths interspacing the nucleosomes. At these ionic strengths ring structures were present, associated with the DNA. At 0.7 M NaCl and above, only DNA filaments were present, occasionally associated with big rings, and nucleosomes were compoetely dissociated. Free DNA molecules were present at all ionic strengths used. The possible origin and significance of the rings are discussed.

Full text

PDF
2315

Images in this article

2317Image
on p.2317
2319Image
on p.2319
2322Image
on p.2322
2323Image
on p.2323
2323Image
on p.2323
2324Image
on p.2324
2324Image
on p.2324
2325Image
on p.2325

Selected References

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

  1. Bakayev V. V., Melnickov A. A., Osicka V. D., Varshausky A. J. Studies on chromatin. II. Isolation and characterization of chromatin subunits. Nucleic Acids Res. 1975 Aug;2(8):1401–1419. doi: 10.1093/nar/2.8.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandt W. F., Von Holt C. Isolation and characterization of the histones from cycad pollen. FEBS Lett. 1975 Mar 1;51(1):84–87. doi: 10.1016/0014-5793(75)80859-3. [DOI] [PubMed] [Google Scholar]
  3. Dubochet J., Ducommun M., Zollinger M., Kellenberger E. A new preparation method for dark-field electron microscopy of biomacromolecules. J Ultrastruct Res. 1971 Apr;35(1):147–167. doi: 10.1016/s0022-5320(71)80148-x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. 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]
  7. 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]
  8. MASTER R. W. POSSIBLE SYNTHESIS OF POLYRIBONUCLEOTIDES OF KNOWN BASE-TRIPLET SEQUENCES. Nature. 1965 Apr 3;206:93–93. doi: 10.1038/206093b0. [DOI] [PubMed] [Google Scholar]
  9. McGhee J. D., Engel J. D. Subunit structure of chromatin is the same in plants and animals. Nature. 1975 Apr 3;254(5499):449–450. doi: 10.1038/254449a0. [DOI] [PubMed] [Google Scholar]
  10. Noll M. Subunit structure of chromatin. Nature. 1974 Sep 20;251(5472):249–251. doi: 10.1038/251249a0. [DOI] [PubMed] [Google Scholar]
  11. Noll M., Thomas J. O., Kornberg R. D. Preparation of native chromatin and damage caused by shearing. Science. 1975 Mar 28;187(4182):1203–1206. doi: 10.1126/science.187.4182.1203. [DOI] [PubMed] [Google Scholar]
  12. Ohlenbusch H. H., Olivera B. M., Tuan D., Davidson N. Selective dissociation of histones from calf thymus nucleoprotein. J Mol Biol. 1967 Apr 28;25(2):299–315. doi: 10.1016/0022-2836(67)90143-x. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Panyim S., Chalkley R. High resolution acrylamide gel electrophoresis of histones. Arch Biochem Biophys. 1969 Mar;130(1):337–346. doi: 10.1016/0003-9861(69)90042-3. [DOI] [PubMed] [Google Scholar]
  16. Paterson R., Knight C. A. Protein synthesis in tobacco protoplasts infected with tobacco mosaic virus. Virology. 1975 Mar;64(1):10–22. doi: 10.1016/0042-6822(75)90074-4. [DOI] [PubMed] [Google Scholar]
  17. Power J. B., Cocking E. C. A simple method for the isolation of very large numbers of leaf protoplasts by using mixtures of cellulase and pectinase. Biochem J. 1969 Mar;111(5):33P–33P. doi: 10.1042/bj1110033pa. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Sperling R., Bustin M. Dynamic equilibrium in histone assembly: self-assembly of single histones and histone pairs. Biochemistry. 1975 Jul 29;14(15):3322–3331. doi: 10.1021/bi00686a006. [DOI] [PubMed] [Google Scholar]
  20. Thomas J. O., Kornberg R. D. An octamer of histones in chromatin and free in solution. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2626–2630. doi: 10.1073/pnas.72.7.2626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Van Holde K. E., Sahasrabuddhe C. G., Shaw B. R. A model for particulate structure in chromatin. Nucleic Acids Res. 1974 Nov;1(11):1579–1586. doi: 10.1093/nar/1.11.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wilhelm X., Champagne M. Dissociation de la nucléoprotéine d'érythrocytes de poulets par les sels. Eur J Biochem. 1969 Aug;10(1):102–109. [PubMed] [Google Scholar]
  23. de Pomerai D. I., Chesterton C. J., Butterworth P. H. Preparation of chromatin. Variation in the template properties of chromatin dependent on the method of perparation. Eur J Biochem. 1974 Aug 1;46(3):461–471. doi: 10.1111/j.1432-1033.1974.tb03639.x. [DOI] [PubMed] [Google Scholar]
  24. 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]

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

RESOURCES