Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1969 Mar;97(3):1134–1141. doi: 10.1128/jb.97.3.1134-1141.1969

Some Properties of Excision-defective Recombination-deficient Mutants of Escherichia coli K-12

Paul Howard-Flanders a, Lee Theriot a, J Brian Stedeford a,1
PMCID: PMC249825  PMID: 4887501

Abstract

Strains of Escherichia coli that carry the mutation uvrA6 show no measurable excision of pyrimidine dimers and are easily killed by ultraviolet (UV) light, whereas strains that carry recA13 are defective in genetic recombination and are also UV-sensitive. An Hfr strain carrying uvrA6 was crossed with an F strain carrying recA13. Among the recombinants identified, one carrying uvrA recA proved to be of exceptional sensitivity to UV light. It is estimated from the UV dose (0.2 erg/mm2 at 253.7 nm) required to reduce the number of colony-forming cells by one natural logarithm that about 1.3 pyrimidine dimers were formed in a genome of 5 × 106 base pairs for each lethal event. This double mutant is 40 times more UV-sensitive than the excision-defective strain carrying uvrA6. The replication of one pyrimidine dimer is generally a lethal event in strains carrying recA13. Spontaneous breakdown and UV-induced breakdown of the deoxyribonucleic acid (DNA) of cells of the various genotypes were estimated by growing the cells in medium containing 3H-thymidine and measuring both acid-precipitable and acid-soluble radioactivity. The UV-induced degradation in strains with recA13 did not require the uvr+ genes and hence appears to depend upon a mechanism other than dimer excision. The greater level of survival after irradiation in Rec+ as compared to Rec bacteria may be due to a recovery mechanism involving the reconstruction of the bacterial chromosome through genetic exchanges which occur between the newly replicated sister duplexes and which effectively circumvent the damaged bases remaining in the DNA.

Full text

PDF
1141

Selected References

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

  1. ADELBERG E. A., BURNS S. N. Genetic variation in the sex factor of Escherichia coli. J Bacteriol. 1960 Mar;79:321–330. doi: 10.1128/jb.79.3.321-330.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BOYCE R. P., HOWARD-FLANDERS P. RELEASE OF ULTRAVIOLET LIGHT-INDUCED THYMINE DIMERS FROM DNA IN E. COLI K-12. Proc Natl Acad Sci U S A. 1964 Feb;51:293–300. doi: 10.1073/pnas.51.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CLARK A. J., MARGULIES A. D. ISOLATION AND CHARACTERIZATION OF RECOMBINATION-DEFICIENT MUTANTS OF ESCHERICHIA COLI K12. Proc Natl Acad Sci U S A. 1965 Feb;53:451–459. doi: 10.1073/pnas.53.2.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark A. J., Chamberlin M., Boyce R. P., Howard-Flanders P. Abnormal metabolic response to ultraviolet light of a recombination deficient mutant of Escherichia coli K12. J Mol Biol. 1966 Aug;19(2):442–454. doi: 10.1016/s0022-2836(66)80015-3. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. FRASER D., JERREL E. A. The amino acid composition of T3 bacteriophage. J Biol Chem. 1953 Nov;205(1):291–295. [PubMed] [Google Scholar]
  7. Fuerst C. R., Siminovitch L. Characterization of an unusual defective lysogenic strain of Escherichia coli K-12(lambda). Virology. 1965 Nov;27(3):449–451. doi: 10.1016/0042-6822(65)90131-5. [DOI] [PubMed] [Google Scholar]
  8. Ganesan A. K., Smith K. C. Recovery of recombination deficient mutants of Escherichia coli K-12 from ultraviolet irradiation. Cold Spring Harb Symp Quant Biol. 1968;33:235–242. doi: 10.1101/sqb.1968.033.01.027. [DOI] [PubMed] [Google Scholar]
  9. Haefner K. Spontaneous lethal sectoring, a further feature of Escherichia coli strains deficient in the function of rec and uvr genes. J Bacteriol. 1968 Sep;96(3):652–659. doi: 10.1128/jb.96.3.652-659.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Howard-Flanders P., Boyce R. P. DNA repair and genetic recombination: studies on mutants of Escherichia coli defective in these processes. Radiat Res. 1966;(Suppl):156+–156+. [PubMed] [Google Scholar]
  11. Howard-Flanders P., Boyce R. P., Theriot L. Three loci in Escherichia coli K-12 that control the excision of pyrimidine dimers and certain other mutagen products from DNA. Genetics. 1966 Jun;53(6):1119–1136. doi: 10.1093/genetics/53.6.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Howard-Flanders P. Genes that control DNA repair and genetic recombination in Escherichia coli. Adv Biol Med Phys. 1968;12:299–317. doi: 10.1016/b978-1-4831-9928-3.50016-3. [DOI] [PubMed] [Google Scholar]
  13. Howard-Flanders P., Rupp W. D., Wilkins B. M., Cole R. S. DNA replication and recombination after UV irradiation. Cold Spring Harb Symp Quant Biol. 1968;33:195–207. doi: 10.1101/sqb.1968.033.01.023. [DOI] [PubMed] [Google Scholar]
  14. Howard-Flanders P., Theriot L. Mutants of Escherichia coli K-12 defective in DNA repair and in genetic recombination. Genetics. 1966 Jun;53(6):1137–1150. doi: 10.1093/genetics/53.6.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Low B. Formation of merodiploids in matings with a class of Rec- recipient strains of Escherichia coli K12. Proc Natl Acad Sci U S A. 1968 May;60(1):160–167. doi: 10.1073/pnas.60.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miura A., Tomizawa J. I. Studies on radiation-sensitive mutants of E. coli. 3. Participation of the rec system in induction of mutation by ultraviolet irradiation. Mol Gen Genet. 1968;103(1):1–10. doi: 10.1007/BF00271151. [DOI] [PubMed] [Google Scholar]
  17. PETTIJOHN D., HANAWALT P. EVIDENCE FOR REPAIR-REPLICATION OF ULTRAVIOLET DAMAGED DNA IN BACTERIA. J Mol Biol. 1964 Aug;9:395–410. doi: 10.1016/s0022-2836(64)80216-3. [DOI] [PubMed] [Google Scholar]
  18. Rupp W. D., Howard-Flanders P. Discontinuities in the DNA synthesized in an excision-defective strain of Escherichia coli following ultraviolet irradiation. J Mol Biol. 1968 Jan 28;31(2):291–304. doi: 10.1016/0022-2836(68)90445-2. [DOI] [PubMed] [Google Scholar]
  19. SETLOW R. B., CARRIER W. L. THE DISAPPEARANCE OF THYMINE DIMERS FROM DNA: AN ERROR-CORRECTING MECHANISM. Proc Natl Acad Sci U S A. 1964 Feb;51:226–231. doi: 10.1073/pnas.51.2.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. SETLOW R. B., SWENSON P. A., CARRIER W. L. THYMINE DIMERS AND INHIBITION OF DNA SYNTHESIS BY ULTRAVIOLET IRRADIATION OF CELLS. Science. 1963 Dec 13;142(3598):1464–1466. doi: 10.1126/science.142.3598.1464. [DOI] [PubMed] [Google Scholar]
  21. Setlow R. B., Carrier W. L., Bollum F. J. Pyrimidine dimers in UV-irradiated poly dI:dC. Proc Natl Acad Sci U S A. 1965 May;53(5):1111–1118. doi: 10.1073/pnas.53.5.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Swenson P. A., Setlow R. B. Effects of ultraviolet radiation on macromolecular synthesis in Escherichia coli. J Mol Biol. 1966 Jan;15(1):201–219. doi: 10.1016/s0022-2836(66)80221-8. [DOI] [PubMed] [Google Scholar]
  23. WULFF D. L. KINETICS OF THYMINE PHOTODIMERIZATION IN DNA. Biophys J. 1963 Sep;3:355–362. doi: 10.1016/s0006-3495(63)86826-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wilkins B. M., Howard-Flanders P. The genetic properties of DNA transferred from ultraviolet-irradiated Hfr cells of Escherichia coli K-12 during mating. Genetics. 1968 Oct;60(2):243–255. doi: 10.1093/genetics/60.2.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Willetts N. S., Clark A. J., Low B. Genetic location of certain mutations conferring recombination deficiency in Escherichia coli. J Bacteriol. 1969 Jan;97(1):244–249. doi: 10.1128/jb.97.1.244-249.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Witkin E. M. Mutation and the repair of radiation damage in bacteria. Radiat Res. 1966;(Suppl):30–53. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES