1887

Abstract

A new procedure has been developed for the isolation of the chromosome complex, termed chromatin, from The bacteria were subjected to low ionic strength and T4 lysozyme, followed by detergent treatment analogous to that employed for the isolation of eukaryotic chromosomes. The chromatin was an insoluble viscous material which contained approximately equal amounts of DNA and RNA. The protein content of the chromatin was almost three times greater than the nucleic acid content. Electron microscopy revealed that the chromatin was highly condensed, having multiple loops and beaded structures with various diameters. The chromatin could be completely solubilized by both micrococcal nuclease and DNAase I, whereas RNAase had no effect. The initial degradation by micrococcal nuclease resulted in the production of a DNA-protein particle, sedimentation coefficient 10S, and an RNA-protein complex of 24S. Further degradation led to a decrease in sedimentation coefficient of the DNA-protein complex, but not of the RNA-protein particle. The peak size of the DNA of the initial DNA-protein particle was approximately 2400 bp. The action of micrococcal nuclease also resulted in the production of several discrete RNA species of various sizes. Several low molecular weight proteins (12000–27000) were found in the DNA-protein complex. The DNA-binding protein HU was present in the undigested chromatin; varying amounts of HU were, however, detected in the DNA-protein and RNA-protein particles.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-128-12-3037
1982-12-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/128/12/mic-128-12-3037.html?itemId=/content/journal/micro/10.1099/00221287-128-12-3037&mimeType=html&fmt=ahah

References

  1. Berthold V., Geider K. 1976; Interaction of DNA with DNA-binding proteins. European Journal of Biochemistry 71:443–449
    [Google Scholar]
  2. Burton K. 1956; A study of conditions and mechanism of the diphenyl reaction of colorimetric estimation of deoxyribonucleic acid. Biochemical Journal 62:315–323
    [Google Scholar]
  3. Griffith J. D. 1976; Visualization of prokaryotic DNA in a regulatory condensed chromatin-like fiber. Proceedings of the National Academy of Sciences of the United States of America 63:563–567
    [Google Scholar]
  4. Hancock R. 1974; Interphase chromosomal deoxy- ribonucleoprotein isolated as a discrete structure from cultured cells. Journal of Molecular Biology 86:649–663
    [Google Scholar]
  5. Hörz W., Altenburger W. 1981; Sequence specific cleavage of DNA by micrococcal nuclease. Nucleic Acids Research 9:2643–2658
    [Google Scholar]
  6. Hübscher U., Lutz H., Kornberg A. 1980; Novel histone H2A-like protein from E. coli. Proceedings of the National Academy of Sciences of the United States of America 77:5097–5101
    [Google Scholar]
  7. Jensen H. B., Kleppe K. 1972; Studies on T4 lysozyme. Affinity for chitin and the use of chitin in the purification of the enzyme. European Journal of Biochemistry 26:305–312
    [Google Scholar]
  8. Jensen H. B., Miron T. 1980; Preparation and properties of insoluble forms of bacteriophage T4 lysozyme and chicken egg white lysozyme. Journal of Solid-Phase Biochemistry 5:45–60
    [Google Scholar]
  9. Jeppesen P. G. 1980; Separation and isolation of DNA fragments using linear polyacrylamide gradient gel electrophoresis. Methods in Enzymology 65:305–319
    [Google Scholar]
  10. Kleppe K., Ohtsuka E., Kleppe R., Molineux I., Khorana H. G. 1971; Studies on polynucleotides. XCVI. Repair replication of short synthetic DNAs as catalyzed by DNA polymerases. Journal of Molecular Biology 56:341–361
    [Google Scholar]
  11. Kleppe K., Øvrebø S., Lossius I. 1979; The bacterial nucleoid. Journal of General Microbiology 112:1–13
    [Google Scholar]
  12. Klug A., Rhodes J., Smith J., Finch J. T., Thomas J. O. 1980; A low resolution structure for the histone core of the nucleosome. Nature; London: 287509–516
    [Google Scholar]
  13. Korch C., Øvrebø S., Kleppe K. 1976; Envelope-associated folded chromosomes from Escherichia coli: variations under diffferent physiological conditions. Journal of Bacteriology 127:904–916
    [Google Scholar]
  14. Kornberg R. D. 1977; Structure of chromatin. Annual Review of Biochemistry 46:931–954
    [Google Scholar]
  15. Laemmli U. K., Favre M. 1973; Maturation of the head of bacteriophage T4. Journal of Molecular Biology 80:575–599
    [Google Scholar]
  16. Laskowski M. 1961; Deoxyribonucleases. In The Enzymes pp. 123–147 Boyer P. D., Lardy H., Myrback K. Edited by New York: Academic Press;
    [Google Scholar]
  17. Maniatis T., Jeffrey A., Van de Sande H. 1975; Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry 14:3787–3794
    [Google Scholar]
  18. Materman E. C., Van Gool A. P. 1978; Compact Escherichia coli nucleoids in a highly supercoiled conformation. Journal of Bacteriology 135:703–706
    [Google Scholar]
  19. Noll M. 1974; Subunit structure of chromatin. Nature; London: 251249–251
    [Google Scholar]
  20. Noll M. 1977; The linkage of chromatin subunits and the role of histone. 1. In Nucleic Acid-Protein Recognition pp. 139–150 Vogel H. J. Edited by New York: Academic Press;
    [Google Scholar]
  21. Pettijohn D. E. 1976; Prokaryotic DNA in nucleoid structure. CRC Critical Reviews in Biochemistry 4:175–202
    [Google Scholar]
  22. Pettijohn D. E., Hecht R. 1973; RNA molecules bound to the folded bacterial genome stabilize DNA folds and segregate domains of supercoiling. Cold Spring Harbor Symposia on Quantitative Biology 38:31–42
    [Google Scholar]
  23. Rouvière-Yaniv J., Gros F. 1975; Characterization of a novel, low-molecular weight DNA-binding protein from Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 72:3428–3432
    [Google Scholar]
  24. Sedmark J. J., Grossberg S. E. . 1977; A rapid, sensitive and versatile assay for protein using Coomassie blue G250. Analytical Biochemistry 79:544–552
    [Google Scholar]
  25. Varshavsky A. J., Newspasov S. A., Bakayev V. V., Bakayeva T. G., Georgiev G. P. 1977; Histone-like proteins in the purified E. coli deoxy-ribonucleoprotein. Nucleic Acids Research 4:2725–2745
    [Google Scholar]
  26. Wallacer R. B., Sargent T. D., Murphy R. F., Bonner J. . 1977; Physical properties of chemically acetylated rat liver chromatin. Proceedings of the National Academy of Sciences of the United States of America 14:3244–3248
    [Google Scholar]
  27. Worcel A., Burgi E. 1972; On the structure of the folded chromosome of Escherichia coli. Journal of Molecular Biology 71:127–147
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-128-12-3037
Loading
/content/journal/micro/10.1099/00221287-128-12-3037
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error