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. 1973 Dec;116(3):1364–1374. doi: 10.1128/jb.116.3.1364-1374.1973

Effect of Rifampin on the Structure and Membrane Attachment of the Nucleoid of Escherichia coli

P Dworsky a,1, M Schaechter a
PMCID: PMC246496  PMID: 4584813

Abstract

Deoxyribonucleic acid (DNA) of Escherichia coli was found to be attached to the cell membrane at about 20 points. This was determined by fractionation of X-irradiated cells with the M band (magnesium-Sarkosyl crystals) technique. The number of attachment points was computed from the relationship between the amount of DNA in M bands and the number of double-strand breaks introduced by the X-ray treatment. The number of attachment points was decreased fourfold by treatment of cells with rifampin. This effect was apparently due to the action of the drug on ribonucleic acid (RNA) polymerase since the drug did not affect a mutant whose RNA polymerase is resistant to rifampin. This suggests that there may be two classes of attachment points of DNA on the membrane, some of which are removed by rifampin treatment and some which are not. Rifampin treatment also resulted in the uncondensing of isolated nucleoids and in an axial appearance of the nucleoids in ultrathin sections. The results suggest that RNA polymerase plays a role, direct or indirect, in maintaining the structure of the bacterial nucleoid and in some of its attachment to the membrane.

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Selected References

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

  1. BURGI E., HERSHEY A. D. A relative molecular weight series derived from the nucleic acid of bacteriophage T2. J Mol Biol. 1961 Aug;3:458–472. doi: 10.1016/s0022-2836(61)80058-2. [DOI] [PubMed] [Google Scholar]
  2. BURGI E., HERSHEY A. D. Sedimentation rate as a measure of molecular weight of DNA. Biophys J. 1963 Jul;3:309–321. doi: 10.1016/s0006-3495(63)86823-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brutlag D., Schekman R., Kornberg A. A possible role for RNA polymerase in the initiation of M13 DNA synthesis. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2826–2829. doi: 10.1073/pnas.68.11.2826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burrell A. D., Feldschreiber P., Dean C. J. DNA-membrane association and the repair of double breaks in x-irradiated Micrococcus radiodurans. Biochim Biophys Acta. 1971 Sep 30;247(1):38–53. doi: 10.1016/0005-2787(71)90805-7. [DOI] [PubMed] [Google Scholar]
  5. CUMMINGS D. J. SEDIMENTATION AND BIOLOGICAL PROPERTIES OF T-PHAGES OF ESCHERICHIA COLI. Virology. 1964 Jul;23:408–418. doi: 10.1016/0042-6822(64)90264-8. [DOI] [PubMed] [Google Scholar]
  6. Cooper S., Helmstetter C. E. Chromosome replication and the division cycle of Escherichia coli B/r. J Mol Biol. 1968 Feb 14;31(3):519–540. doi: 10.1016/0022-2836(68)90425-7. [DOI] [PubMed] [Google Scholar]
  7. Fielding P., Fox C. F. Evidence for stable attachment of DNA to membrane at the replication origin of Escherichia coli. Biochem Biophys Res Commun. 1970 Oct 9;41(1):157–162. doi: 10.1016/0006-291x(70)90482-1. [DOI] [PubMed] [Google Scholar]
  8. Fuchs E., Hanawalt P. Isolation and characterization of the DNA replication complex from Escherichia coli. J Mol Biol. 1970 Sep 14;52(2):301–322. doi: 10.1016/0022-2836(70)90032-x. [DOI] [PubMed] [Google Scholar]
  9. Gesteland R. F. Isolation and characterization of ribonuclease I mutants of Escherichia coli. J Mol Biol. 1966 Mar;16(1):67–84. doi: 10.1016/s0022-2836(66)80263-2. [DOI] [PubMed] [Google Scholar]
  10. HERSHEY A. D., CHASE M. Independent functions of viral protein and nucleic acid in growth of bacteriophage. J Gen Physiol. 1952 May;36(1):39–56. doi: 10.1085/jgp.36.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. HERSHEY A. D., GOLDBERG E., BURGI E., INGRAHAM L. Local denaturation of DNA by shearing forces and by heat. J Mol Biol. 1963 Mar;6:230–243. doi: 10.1016/s0022-2836(63)80072-8. [DOI] [PubMed] [Google Scholar]
  12. Helmstetter C. E. DNA synthesis during the division cycle of rapidly growing Escherichia coli B/r. J Mol Biol. 1968 Feb 14;31(3):507–518. doi: 10.1016/0022-2836(68)90424-5. [DOI] [PubMed] [Google Scholar]
  13. Ivarie R. D., Pène J. J. Association of many regions of the Bacillus subtilis chromosome with the cell membrane. J Bacteriol. 1973 May;114(2):571–576. doi: 10.1128/jb.114.2.571-576.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ivarie R. D., Pène J. J. Association of the Bacillus subtilis chromosome with the cell membrane: resolution of free and bound deoxyribonucleic acid on renografin gradients. J Bacteriol. 1970 Nov;104(2):839–850. doi: 10.1128/jb.104.2.839-850.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. KELLENBERGER E., RYTER A., SECHAUD J. Electron microscope study of DNA-containing plasms. II. Vegetative and mature phage DNA as compared with normal bacterial nucleoids in different physiological states. J Biophys Biochem Cytol. 1958 Nov 25;4(6):671–678. doi: 10.1083/jcb.4.6.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Levin D., Hutchinson F. Neutral sucrose sedimentation of very large DNA from Bacillus subtilis. I. Effect of random double-strand breaks and centrifuge speed on sedimentation. J Mol Biol. 1973 Apr 15;75(3):455–478. doi: 10.1016/0022-2836(73)90454-3. [DOI] [PubMed] [Google Scholar]
  18. Morgan C., Rosenkranz H. S., Carr H. S., Rose H. M. Electron microscopy of chloramphenicol-treated Escherichia coli. J Bacteriol. 1967 Jun;93(6):1987–2002. doi: 10.1128/jb.93.6.1987-2002.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ormerod M. G., Lehmann A. R. The release of high molecular weight DNA from a mammalian cell (L-5178Y). Attachment of the DNA to the nuclear membrane. Biochim Biophys Acta. 1971 Jan 28;228(2):331–343. [PubMed] [Google Scholar]
  20. Pyeritz R. E., Schlegel R. A., Thomas C. A., Jr Hydrodynamic shear breakage of DNA may produce single-chained terminals. Biochim Biophys Acta. 1972 Jul 31;272(4):504–509. doi: 10.1016/0005-2787(72)90505-9. [DOI] [PubMed] [Google Scholar]
  21. RYTER A. [Electron microscopic study of the nuclear transformations 05 E. coli K12S and K12S (lambda 26) after irradiation with ultraviolet rays and x-rays]. J Biophys Biochem Cytol. 1960 Oct;8:399–412. [PMC free article] [PubMed] [Google Scholar]
  22. Rosenberg B. H., Cavalieri L. F. Shear and the melting of DNA: an especially sensitive portion of the E. coli genome. Biophys J. 1968 Oct;8(10):1138–1145. doi: 10.1016/s0006-3495(68)86545-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rosenberg B. H., Cavalieri L. F. Shear sensitivity of the E. coli genome: multiple membrane attachment points of the E. coli DNA. Cold Spring Harb Symp Quant Biol. 1968;33:65–72. doi: 10.1101/sqb.1968.033.01.012. [DOI] [PubMed] [Google Scholar]
  24. Ryter A. Association of the nucleus and the membrane of bacteria: a morphological study. Bacteriol Rev. 1968 Mar;32(1):39–54. doi: 10.1128/br.32.1.39-54.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ryter A., Schaeffer P., Ionesco H. Classification cytologique, par leur stade de blocage, des mutants de sporulation de Bacillus subtilis Marburg. Ann Inst Pasteur (Paris) 1966 Mar;110(3):305–315. [PubMed] [Google Scholar]
  26. Sadowski P. D., Kerr C. Degradation of Escherichia coli B deoxyribonucleic acid after infection with deoxyribonucleic acid-defective amber mutants of bacteriophage T7. J Virol. 1970 Aug;6(2):149–155. doi: 10.1128/jvi.6.2.149-155.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shull F. W., Jr, Fralick J. A., Stratton L. P., Fisher W. D. Membrane association of conjugally transferred deoxyribonucleic acid in Escherichia coli minicells. J Bacteriol. 1971 May;106(2):626–633. doi: 10.1128/jb.106.2.626-633.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smith D. W., Hanawalt P. C. Properties of the growing point region in the bacterial chromosome. Biochim Biophys Acta. 1967 Dec 19;149(2):519–531. doi: 10.1016/0005-2787(67)90180-3. [DOI] [PubMed] [Google Scholar]
  29. Stonington O. G., Pettijohn D. E. The folded genome of Escherichia coli isolated in a protein-DNA-RNA complex. Proc Natl Acad Sci U S A. 1971 Jan;68(1):6–9. doi: 10.1073/pnas.68.1.6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sueoka N., Quinn W. G. Membrane attachment of the chromosome replication origin in Bacillus subtilis. Cold Spring Harb Symp Quant Biol. 1968;33:695–705. doi: 10.1101/sqb.1968.033.01.078. [DOI] [PubMed] [Google Scholar]
  31. Tremblay G. Y., Daniels M. J., Schaechter M. Isolation of a cell membrane-DNA-nascent RNA complex from bacteria. J Mol Biol. 1969 Feb 28;40(1):65–76. doi: 10.1016/0022-2836(69)90296-4. [DOI] [PubMed] [Google Scholar]
  32. Worcel A., Burgi E. On the structure of the folded chromosome of Escherichia coli. J Mol Biol. 1972 Nov 14;71(2):127–147. doi: 10.1016/0022-2836(72)90342-7. [DOI] [PubMed] [Google Scholar]

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