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. 1972 Apr;110(1):71–80. doi: 10.1128/jb.110.1.71-80.1972

Photoreactivation, Excision, and Strand-Rejoining Repair in R Factor-Containing Minicells of Escherchia coli K-12

M C Paterson 1, K J Roozen 1
PMCID: PMC247380  PMID: 4553007

Abstract

Chromosomeless “minicells” are formed by misplaced cell fissions near the polar extremities of an Escherichia coli K-12 mutant strain. Resistance (R)-factor deoxyribonucleic acid (DNA) can be introduced into minicells by segregation from an R+ (R64-11) derivative of the original mutant. We have assessed the ability of R+ minicells to correct defects produced in their plasmid DNA by ultraviolet (UV) and gamma radiations. Minicells harboring plasmid DNA, in comparison with their repair-proficient minicell-producing parents, possess (i) an equal competence to rejoin single-strand breaks induced in DNA by gamma rays, (ii) a reduced capacity for the photoenzymatic repair of UV-induced pyrimidine dimers, and (iii) a total inability to excise dimers, apparently owing to a deficiency in UV-specific endonuclease activity responsible for mediating the initial incision step in excision repair. Assuming that the DNA repair properties of R+ minicells reflect the concentration of repair enzymes located in the plasmid-containing polar caps of entire cells, these findings suggest that: (i) the enzymes responsible for rejoining single-strand breaks are distributed throughout the cell; (ii) photoreactivating enzyme molecules tend to be concentrated near bacterial DNA and to a lesser extent near plasmid DNA; and (iii) UV-specific endonuclease molecules are primarily confined to the central region of the E. coli cell and, thus, seldom segregate with R-factor DNA into minicells.

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Selected References

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

  1. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axelrod D. E., Adler H. I. Influence of the fertility episome on the survival of x-irradiated Escherichia coli. J Bacteriol. 1969 Apr;98(1):329–330. doi: 10.1128/jb.98.1.329-330.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyle J. M., Setlow R. B. Correlations between host-cell reactivation, ultraviolet reactivation and pyrimidine dimer excision in the DNA of bacteriophage lambda. J Mol Biol. 1970 Jul 14;51(1):131–144. doi: 10.1016/0022-2836(70)90275-5. [DOI] [PubMed] [Google Scholar]
  4. Carrier W. L., Setlow R. B. Paper strip method for assaying gradient fractions containing radioactive macromolecules. Anal Biochem. 1971 Oct;43(2):427–432. doi: 10.1016/0003-2697(71)90272-7. [DOI] [PubMed] [Google Scholar]
  5. Cohen A., Fisher W. D., Curtiss R., 3rd, Adler H. I. The properties of DNA transferred to minicells during conjugation. Cold Spring Harb Symp Quant Biol. 1968;33:635–641. doi: 10.1101/sqb.1968.033.01.071. [DOI] [PubMed] [Google Scholar]
  6. Fraser D., Mahler H. R., Shug A. L., Thomas C. A. THE INFECTION OF SUB-CELLULAR ESCHERICHIA COLI, STRAIN B, WITH A DNA PREPARATION FROM T2 BACTERIOPHAGE. Proc Natl Acad Sci U S A. 1957 Nov 15;43(11):939–947. doi: 10.1073/pnas.43.11.939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Freifelder D., Folkmanis A., Kirschner I. Studies on Escherichia coli sex factors: evidence that covalent circles exist within cells and the general problem of isolation of covalent circles. J Bacteriol. 1971 Mar;105(3):722–727. doi: 10.1128/jb.105.3.722-727.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Freifelder D. Rate of production of single-strand breaks in DNA by x-irradiation in situ. J Mol Biol. 1968 Jul 28;35(2):303–309. doi: 10.1016/s0022-2836(68)80026-9. [DOI] [PubMed] [Google Scholar]
  9. Grossman L., Kaplan J. C., Kushner S. R., Mahler I. Enzymes involved in the early stages of repair of ultraviolet-irradiated DNA. Cold Spring Harb Symp Quant Biol. 1968;33:229–234. doi: 10.1101/sqb.1968.033.01.026. [DOI] [PubMed] [Google Scholar]
  10. Howard-Flanders P. DNA repair. Annu Rev Biochem. 1968;37:175–200. doi: 10.1146/annurev.bi.37.070168.001135. [DOI] [PubMed] [Google Scholar]
  11. Howarth S. Resistance to the bactericidal effect of ultraviolet radiation conferred on Enterobacteria by the colicine factor coli. J Gen Microbiol. 1965 Jul;40(1):43–55. doi: 10.1099/00221287-40-1-43. [DOI] [PubMed] [Google Scholar]
  12. Inselburg J. Segregation into and replication of plasmid deoxyribonucleic acid in chromosomeless segregants of Escherichia coli. J Bacteriol. 1970 Jun;102(3):642–647. doi: 10.1128/jb.102.3.642-647.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kass L. R., Yarmolinsky M. B. Segregation of functional sex factor into minicells. Proc Natl Acad Sci U S A. 1970 Jul;66(3):815–822. doi: 10.1073/pnas.66.3.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Levy S. B., Norman P. Segregation of transferable R factors into Escherichia coli minicells. Nature. 1970 Aug 8;227(5258):606–607. doi: 10.1038/227606a0. [DOI] [PubMed] [Google Scholar]
  15. McGrath R. A., Williams R. W. Reconstruction in vivo of irradiated Escherichia coli deoxyribonucleic acid; the rejoining of broken pieces. Nature. 1966 Oct 29;212(5061):534–535. doi: 10.1038/212534a0. [DOI] [PubMed] [Google Scholar]
  16. Paterson M. C., Boyle J. M., Setlow R. B. Ultraviolet- and X-ray-induced responses of a deoxyribonucleic acid polymerase-deficient mutant of Escherichia coli. J Bacteriol. 1971 Jul;107(1):61–67. doi: 10.1128/jb.107.1.61-67.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Roozen K. J., Fenwick R. G., Jr, Curtiss R., 3rd Synthesis of ribonucleic acid and protein in plasmid-containing minicells of Escherichia coli K-12. J Bacteriol. 1971 Jul;107(1):21–33. doi: 10.1128/jb.107.1.21-33.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. STUDIER F. W. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA. J Mol Biol. 1965 Feb;11:373–390. doi: 10.1016/s0022-2836(65)80064-x. [DOI] [PubMed] [Google Scholar]
  19. Town C. D., Smith K. C., Kaplan H. S. DNA polymerase required for rapid repair of x-ray--induced DNA strand breaks in vivo. Science. 1971 May 21;172(3985):851–854. doi: 10.1126/science.172.3985.851. [DOI] [PubMed] [Google Scholar]
  20. Vapnek D., Lipman M. B., Rupp W. D. Physical properties and mechanism of transfer of R factors in Escherichia coli. J Bacteriol. 1971 Oct;108(1):508–514. doi: 10.1128/jb.108.1.508-514.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

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