Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1968 Oct;8(10):1119–1130. doi: 10.1016/S0006-3495(68)86543-9

Recovery of Colony-Forming Ability and Genetic Marker Activity by UV-Damaged Hemophilus influenzae

Rufus S Day III, R A Deering
PMCID: PMC1367659  PMID: 5302986

Abstract

The rate of recovery of UV-irradiated Hemophilus influenzae from acriflavine-sensitized loss of colony-forming ability was studied at various acriflavine concentrations, UV doses, and temperatures. This rate (as calculated from an equation based upon certain assumptions) was on the order of 0.07 per minute per cell at 37°C. This did not vary greatly with UV dose or acriflavine concentration, but did with temperature, giving a ΔH‡ of about 16 kcal/mole. In another set of experiments, cells bearing two genetic markers (resistance to 2000 μg/ml streptomycin and to 2.5 μg/ml novobiocin) were irradiated and then incubated without acriflavine. DNA extracts made from samples taken after various periods of incubation time were assayed on antibiotic-sensitive cells using acriflavine to inhibit repair during and following transformation. It was found that both in vivo irradiated markers were reactivated in the donor to approximately the same extent (with a rate constant of 0.04 per minute). This result was in contrast to the results obtained when extracted DNA bearing the same markers was irradiated in vitro and used to transform cells. In this latter case the streptomycin marker was much more sensitive than the novobiocin marker. This difference is interpreted as being due to the mechanics of the transformation system.

Full text

PDF
1119

Selected References

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

  1. ALPER T. EFFECTS ON IRRADIATED MICRO-ORGANISMS OF GROWTH IN THE PRESENCE OF ACRIFLAVINE. Nature. 1963 Nov 9;200:534–536. doi: 10.1038/200534a0. [DOI] [PubMed] [Google Scholar]
  2. BARNHART B. J., HERRIOTT R. M. PENETRATION OF DEOXYRIBONUCLEIC ACID INTO HEMOPHILUS INFLUENZAE. Biochim Biophys Acta. 1963 Sep 17;76:25–39. [PubMed] [Google Scholar]
  3. 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]
  4. Bresler S. E., Kalinin V. L., Perumov D. A. Inactivation and mutagenesis on isolated DNA. I. Theory of inactivation of transforming DNA. Mutat Res. 1967 Jul-Aug;4(4):389–398. doi: 10.1016/0027-5107(67)90001-2. [DOI] [PubMed] [Google Scholar]
  5. Doudney C. O., White B. F., Bruce B. J. Acriflavine modification of nucleic acid formation, mutation induction and survival in ultraviolet light exposed bacteria. Biochem Biophys Res Commun. 1964 Feb 18;15(1):70–75. doi: 10.1016/0006-291x(64)90105-6. [DOI] [PubMed] [Google Scholar]
  6. FEINER R. R., HILL R. F. EFFECT OF BASIC DYES ON HOST-CELL REACTIVATION OF ULTRA-VIOLETIRRADIATED PHAGE. Nature. 1963 Oct 19;200:291–293. doi: 10.1038/200291a0. [DOI] [PubMed] [Google Scholar]
  7. Harm W. Differential effects of acriflavine and caffeine on various ultraviolet-irradiated Escherichia coli strains and T1 phage. Mutat Res. 1967 Mar-Apr;4(2):93–110. doi: 10.1016/0027-5107(67)90061-9. [DOI] [PubMed] [Google Scholar]
  8. Harm W. The role of host-cell repair in liquid-holding recovery of u.v.-irradiated Escherichia coli. Photochem Photobiol. 1966 Sep;5(9):747–760. doi: 10.1111/j.1751-1097.1966.tb05824.x. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Mahler I. Characteristics of an ultraviolet irradiation sensitive strain of Bacillus subtilis. Biochem Biophys Res Commun. 1965 Nov 22;21(4):384–391. doi: 10.1016/0006-291x(65)90206-8. [DOI] [PubMed] [Google Scholar]
  11. Notani N., Goodgal S. H. On the nature of recombinants formed during transformation in Hemophilus influenzae. J Gen Physiol. 1966 Jul;49(6):197–209. doi: 10.1085/jgp.49.6.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Okubo S., Romig W. R. Comparison of ultraviolet sensitivity of Bacillus subtilis bacteriophage SPO2 and its infectious DNA. J Mol Biol. 1965 Nov;14(1):130–142. doi: 10.1016/s0022-2836(65)80235-2. [DOI] [PubMed] [Google Scholar]
  13. Patrick M. H., Rupert C. S. The effects of host-cell reactivation on assay of U.V.-irradiated Haemophilus influenzae transforming DNA. Photochem Photobiol. 1967 Jan;6(1):1–20. doi: 10.1111/j.1751-1097.1967.tb08785.x. [DOI] [PubMed] [Google Scholar]
  14. Reiter H., Strauss B. Repair of damage induced by a monofunctional alkylating agent in a transformable, ultraviolet-sensitive strain of Bacillus subtilis. J Mol Biol. 1965 Nov;14(1):179–194. doi: 10.1016/s0022-2836(65)80239-x. [DOI] [PubMed] [Google Scholar]
  15. Roberts R. B., Aldous E. RECOVERY FROM ULTRAVIOLET IRRADIATION IN ESCHERICHIA COLI. J Bacteriol. 1949 Mar;57(3):363–375. doi: 10.1128/jb.57.3.363-375.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rupert C. S. Shapes of the U.V. inactivation curves for single and linked double markers in Haemophilus influenzae transforming DNA. Photochem Photobiol. 1968 May;7(5):437–449. doi: 10.1111/j.1751-1097.1968.tb07405.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. SETLOW R. B. PHYSICAL CHANGES AND MUTAGENESIS. J Cell Physiol. 1964 Oct;64:SUPPL 1–1:68. [PubMed] [Google Scholar]
  19. STUY J. H. Studies on the radiation inactivation of microorganisms. V. Deoxyribonucleic acid metabolism in ultraviolet-irradiated Haemophilus influenzae. J Bacteriol. 1959 Jul;78(1):49–58. doi: 10.1128/jb.78.1.49-58.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Setlow R. B., Carrier W. L. Pyrimidine dimers in ultraviolet-irradiated DNA's. J Mol Biol. 1966 May;17(1):237–254. doi: 10.1016/s0022-2836(66)80105-5. [DOI] [PubMed] [Google Scholar]
  21. Voll M. J., Goodgal S. H. Loss of activity of transforming deoxyribonucleic acid after uptake by Haemophilus influenzae. J Bacteriol. 1965 Oct;90(4):873–883. doi: 10.1128/jb.90.4.873-883.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. WITKIN E. M. THE EFFECT OF ACRIFLAVINE ON PHOTOREVERSAL OF LETHAL AND MUTAGENIC DAMAGE PRODUCED IN BACTERIA BY ULTRAVIOLET LIGHT. Proc Natl Acad Sci U S A. 1963 Sep;50:425–430. doi: 10.1073/pnas.50.3.425. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES