Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1976 Feb;125(2):643–654. doi: 10.1128/jb.125.2.643-654.1976

Lethal effect of mitomycin C on Haemophilus influenzae.

G D Small, J K Setlow, J Kooistra, R Shapanka
PMCID: PMC236126  PMID: 173712

Abstract

The sensitivity of ultraviolet-sensitive strains to inactivation by mitomycin C (MC) is at the most only a factor of two greater than that of the wild type. The presence of inducible prophage has very little effect on the sensitivity. Genes which control excision of ultraviolet-induced pyrimidine dimers also control repair of MC-induced cross-links, as measured by resistance of denatured deoxyribonucleic acid (DNA) from treated cells to S1 nuclease digestion. However, endonucleolytic breaks in MC-damaged DNA, as judged by decreased single-strand molecular weight upon incubation of treated cells, are independent of these genes and probably are caused by monoadducts. After long periods of incubation there is a return to the molecular weight of untreated DNA. DNA degradation after MC treatment of various strains is not correlated with sensitivity to inactivation. Stationary-phase cells of all strains are more than twice as sensitive to MC as exponentially growing cells, and the sensitivity difference agrees with the measured difference in the number of cross-links after MC treatment of cells in the two growth stages. Evidence has been obtained that these phenomena result from differences in uptake of MC, which can be influenced by cyclic adenosine monophosphate. Small deviations in MC sensitivity from that of the wild type observed in mutants lacking the adenosine 5'-triphosphate-dependent nuclease are postulated to result from differences in MC uptake. These mutants, although no more ultraviolet sensitive than the wild type, are more sensitive to streptomycin, which also must be taken up by the cell to be effective.

Full text

PDF
643

Selected References

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

  1. Barnhart B. J., Cox S. H. Radiation-sensitive and radiation-resistant mutants of Haemophilus influenzae. J Bacteriol. 1968 Jul;96(1):280–282. doi: 10.1128/jb.96.1.280-282.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boling M. E., Allison D. P., Setlow J. K. Bacteriophage of Haemophilus influenzae. 3. Morphology, DNA homology, and immunity properties of HPlcl, S2, and the defective bacteriophage from strain Rd. J Virol. 1973 Apr;11(4):585–591. doi: 10.1128/jvi.11.4.585-591.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boling M. E., Setlow J. K. Dependence of Vegetative Recombination Among Haemophilus influenzae Bacteriophage on the Host Cell. J Virol. 1969 Sep;4(3):240–243. doi: 10.1128/jvi.4.3.240-243.1969. [DOI] [PMC free article] [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. Cole R. S. Inactivation of Escherichia coli, F' episomes at transfer, and bacteriophage lambda by psoralen plus 360-nm light: significance of deoxyribonucleic acid cross-links. J Bacteriol. 1971 Sep;107(3):846–852. doi: 10.1128/jb.107.3.846-852.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cole R. S. Repair of DNA containing interstrand crosslinks in Escherichia coli: sequential excision and recombination. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1064–1068. doi: 10.1073/pnas.70.4.1064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Friedman E. A., Smith H. O. An adenosine triphosphate-dependent deoxyribonuclease from Hemophilus influenzae Rd. I. Purification and properties of the enzyme. J Biol Chem. 1972 May 10;247(9):2846–2853. [PubMed] [Google Scholar]
  8. Gillis M., De Ley J., De Cleene M. The determination of molecular weight of bacterial genome DNA from renaturation rates. Eur J Biochem. 1970 Jan;12(1):143–153. doi: 10.1111/j.1432-1033.1970.tb00831.x. [DOI] [PubMed] [Google Scholar]
  9. IIJIMA T., HAGIWARA A. Mutagenic action of mitomycin C on Escherichia coli. Nature. 1960 Feb 6;185:395–396. doi: 10.1038/185395b0. [DOI] [PubMed] [Google Scholar]
  10. IYER V. N., SZYBALSKI W. A MOLECULAR MECHANISM OF MITOMYCIN ACTION: LINKING OF COMPLEMENTARY DNA STRANDS. Proc Natl Acad Sci U S A. 1963 Aug;50:355–362. doi: 10.1073/pnas.50.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. IYER V. N., SZYBALSKI W. MITOMYCINS AND PORFIROMYCIN: CHEMICAL MECHANISM OF ACTIVATION AND CROSS-LINKING OF DNA. Science. 1964 Jul 3;145(3627):55–58. doi: 10.1126/science.145.3627.55. [DOI] [PubMed] [Google Scholar]
  12. Kooistra J., Venema G. Fate of donor DNA in some poorly transformable strains of Haemophilus influenzae. Mutat Res. 1970 Mar;9(3):245–253. doi: 10.1016/0027-5107(70)90126-0. [DOI] [PubMed] [Google Scholar]
  13. Kooistra J., Venema G. Fate of donor deoxyribonucleic acid in a highly transformation-deficient strain of Haemophilus influenzae. J Bacteriol. 1974 Sep;119(3):705–717. doi: 10.1128/jb.119.3.705-717.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kooistra J., Venema G. Poor transformability with novr and eryr donor DNA of some mitomycin-C-sensitive strains of Haemophilus influenzae. Mutat Res. 1973 Dec;20(3):313–326. doi: 10.1016/0027-5107(73)90054-7. [DOI] [PubMed] [Google Scholar]
  15. LeClerc J. E., Setlow J. K. Effects of combining ultraviolet repair and recombination mutations in Haemophilus influenzae. Nat New Biol. 1973 Feb 7;241(110):172–174. doi: 10.1038/newbio241172a0. [DOI] [PubMed] [Google Scholar]
  16. Leclerc J. E., Setlow J. K. Postreplication repair of ultraviolet damage in Haemophilus influenzae. J Bacteriol. 1972 Jun;110(3):930–934. doi: 10.1128/jb.110.3.930-934.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mahler I. Effect of mitomycin C on five excision-repair mutants of Bacillus subtilis. Biochem Biophys Res Commun. 1966 Oct 5;25(1):73–79. doi: 10.1016/0006-291x(66)90642-5. [DOI] [PubMed] [Google Scholar]
  18. Modak S. P., Setlow J. K. Synthesis of deoxyribonucleic acid after ultraviolet irradiation of sensitive and resistant Haemophilus influenzae. J Bacteriol. 1969 Jun;98(3):1195–1198. doi: 10.1128/jb.98.3.1195-1198.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. OTSUJI N., SEKIGUCHI M., IIJIMA T., TAKAGI Y. Induction of phage formation in the lysogenic Escherichia coli K-12 by mitomycin C. Nature. 1959 Oct 3;184(Suppl 14):1079–1080. doi: 10.1038/1841079b0. [DOI] [PubMed] [Google Scholar]
  20. Okubo S., Romig W. R. Impaired transformability of Bacillus subtilis mutant sensitive to mitomycin C and ultraviolet radiation. J Mol Biol. 1966 Feb;15(2):440–454. doi: 10.1016/s0022-2836(66)80120-1. [DOI] [PubMed] [Google Scholar]
  21. Setlow J. K., Boling M. E., Allison D. P., Beattie K. L. Relationship between prophage induction and transformation in Haemophilus influenzae. J Bacteriol. 1973 Jul;115(1):153–161. doi: 10.1128/jb.115.1.153-161.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Setlow J. K., Boling M. E., Beattie K. L., Kimball R. F. A complex of recombination and repair genes in Haemophilus influenzae. J Mol Biol. 1972 Jul 21;68(2):361–378. doi: 10.1016/0022-2836(72)90218-5. [DOI] [PubMed] [Google Scholar]
  23. Setlow J. K., Brown D. C., Boling M. E., Mattingly A., Gordon M. P. Repair of deoxyribonucleic acid in Haemophilus influenzae. I. X-ray sensitivity of ultraviolet-sensitive mutants and their behavior as hosts to ultraviolet-irradiated bacteriophage and transforming deoxyribonucleic acid. J Bacteriol. 1968 Feb;95(2):546–558. doi: 10.1128/jb.95.2.546-558.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sutton W. D. A crude nuclease preparation suitable for use in DNA reassociation experiments. Biochim Biophys Acta. 1971 Jul 29;240(4):522–531. doi: 10.1016/0005-2787(71)90709-x. [DOI] [PubMed] [Google Scholar]
  25. Terawaki A., Greenberg J. Post-treatment breakage of mitomycin C induced cross-links in deoxyribonucleic acid of Escherichia coli. Biochim Biophys Acta. 1966 Jun 22;119(3):540–546. doi: 10.1016/0005-2787(66)90130-4. [DOI] [PubMed] [Google Scholar]
  26. Tomasz M., Mercado C. M., Olson J., Chatterjie N. The mode of interaction of mitomycin C with deoxyribonucleic acid and other polynucleotides in vitro. Biochemistry. 1974 Nov 19;13(24):4878–4887. doi: 10.1021/bi00721a002. [DOI] [PubMed] [Google Scholar]
  27. Wilcox K. W., Smith H. O. Isolation and characterization of mutants of Haemophilus influenzae deficient in an adenosine 5'-triphosphate-dependent deoxyribonuclease activity. J Bacteriol. 1975 May;122(2):443–453. doi: 10.1128/jb.122.2.443-453.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wise E. M., Jr, Alexander S. P., Powers M. Adenosine 3':5'-cyclic monophosphate as a regulator of bacterial transformation. Proc Natl Acad Sci U S A. 1973 Feb;70(2):471–474. doi: 10.1073/pnas.70.2.471. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES