Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Jan;161(1):123–127. doi: 10.1128/jb.161.1.123-127.1985

Isolation and characterization of an Escherichia coli mutant lacking the cytochrome o terminal oxidase.

D C Au, R M Lorence, R B Gennis
PMCID: PMC214844  PMID: 2981797

Abstract

A respiration-deficient mutant of Escherichia coli has been isolated which is unable to grow aerobically on nonfermentable substrates such as succinate and lactate. Spectroscopic and immunological studies showed that this mutant lacks the cytochrome o terminal oxidase of the high aeration branch of the aerobic electron transport chain. This strain carries a mutation in a gene designated cyo which is cotransducible with the acrA locus. Mutations in cyo were obtained by mutagenizing a strain that was cyd and, thus, was lacking the cytochrome d terminal oxidase. Strain RG99, which carries both the cyd- and cyo- alleles, grows normally under anaerobic conditions in the presence of nitrate. Introduction of the cyd+ allele into the strain restores the respiration function of the strain, indicating that the cytochrome o branch of the respiratory chain is dispensable under normal laboratory growth conditions.

Full text

PDF
124

Images in this article

Selected References

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

  1. Au D. C., Green G. N., Gennis R. B. Role of quinones in the branch of the Escherichia coli respiratory chain that terminates in cytochrome o. J Bacteriol. 1984 Jan;157(1):122–125. doi: 10.1128/jb.157.1.122-125.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Green G. N., Gennis R. B. Isolation and characterization of an Escherichia coli mutant lacking cytochrome d terminal oxidase. J Bacteriol. 1983 Jun;154(3):1269–1275. doi: 10.1128/jb.154.3.1269-1275.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hackett N. R., Bragg P. D. Membrane cytochromes of Escherichia coli grown aerobically and anaerobically with nitrate. J Bacteriol. 1983 May;154(2):708–718. doi: 10.1128/jb.154.2.708-718.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hendler R. W., Shrager R. I. Potentiometric analysis of Escherichia coli cytochromes in the optical absorbance range of 500 nm to 700 nm. J Biol Chem. 1979 Nov 25;254(22):11288–11299. [PubMed] [Google Scholar]
  7. Hendler R. W., Towne D. W., Shrager R. I. Redox properties of beta-type cytochromes in Escherichia coli and rat liver mitochondria and techniques for their analysis. Biochim Biophys Acta. 1975 Jan 31;376(1):42–62. doi: 10.1016/0005-2728(75)90203-0. [DOI] [PubMed] [Google Scholar]
  8. Kita K., Kasahara M., Anraku Y. Formation of a membrane potential by reconstructed liposomes made with cytochrome b562-o complex, a terminal oxidase of Escherichia coli K12. J Biol Chem. 1982 Jul 25;257(14):7933–7935. [PubMed] [Google Scholar]
  9. Kita K., Konishi K., Anraku Y. Terminal oxidases of Escherichia coli aerobic respiratory chain. I. Purification and properties of cytochrome b562-o complex from cells in the early exponential phase of aerobic growth. J Biol Chem. 1984 Mar 10;259(5):3368–3374. [PubMed] [Google Scholar]
  10. Kita K., Konishi K., Anraku Y. Terminal oxidases of Escherichia coli aerobic respiratory chain. II. Purification and properties of cytochrome b558-d complex from cells grown with limited oxygen and evidence of branched electron-carrying systems. J Biol Chem. 1984 Mar 10;259(5):3375–3381. [PubMed] [Google Scholar]
  11. Kita K., Yamato I., Anraku Y. Purification and properties of cytochrome b556 in the respiratory chain of aerobically grown Escherichia coli K12. J Biol Chem. 1978 Dec 25;253(24):8910–8915. [PubMed] [Google Scholar]
  12. Koland J. G., Miller M. J., Gennis R. B. Reconstitution of the membrane-bound, ubiquinone-dependent pyruvate oxidase respiratory chain of Escherichia coli with the cytochrome d terminal oxidase. Biochemistry. 1984 Jan 31;23(3):445–453. doi: 10.1021/bi00298a008. [DOI] [PubMed] [Google Scholar]
  13. Kranz R. G., Barassi C. A., Miller M. J., Green G. N., Gennis R. B. Immunological characterization of an Escherichia coli strain which is lacking cytochrome d. J Bacteriol. 1983 Oct;156(1):115–121. doi: 10.1128/jb.156.1.115-121.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kranz R. G., Gennis R. B. Immunological characterization of the cytochrome o terminal oxidase from Escherichia coli. J Biol Chem. 1983 Sep 10;258(17):10614–10621. [PubMed] [Google Scholar]
  15. Lorence R. M., Green G. N., Gennis R. B. Potentiometric analysis of the cytochromes of an Escherichia coli mutant strain lacking the cytochrome d terminal oxidase complex. J Bacteriol. 1984 Jan;157(1):115–121. doi: 10.1128/jb.157.1.115-121.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Matsushita K., Patel L., Gennis R. B., Kaback H. R. Reconstitution of active transport in proteoliposomes containing cytochrome o oxidase and lac carrier protein purified from Escherichia coli. Proc Natl Acad Sci U S A. 1983 Aug;80(16):4889–4893. doi: 10.1073/pnas.80.16.4889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller M. J., Gennis R. B. The purification and characterization of the cytochrome d terminal oxidase complex of the Escherichia coli aerobic respiratory chain. J Biol Chem. 1983 Aug 10;258(15):9159–9165. [PubMed] [Google Scholar]
  19. Poole R. K., Chance B. The reaction of cytochrome o in Escherichia coli K12 with oxygen. Evidence for a spectrally and kinetically distinct cytochrome o in cells from oxygen-limited cultures. J Gen Microbiol. 1981 Oct;126(2):277–287. doi: 10.1099/00221287-126-2-277. [DOI] [PubMed] [Google Scholar]
  20. Pudek M. R., Bragg P. D. Redox potentials of the cytochromes in the respiratory chain of aerobically grown Escherichia coli. Arch Biochem Biophys. 1976 Jun;174(2):546–552. doi: 10.1016/0003-9861(76)90382-9. [DOI] [PubMed] [Google Scholar]
  21. Rice C. W., Hempfling W. P. Oxygen-limited continuous culture and respiratory energy conservation in Escherichia coli. J Bacteriol. 1978 Apr;134(1):115–124. doi: 10.1128/jb.134.1.115-124.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shipp W. S. Cytochromes of Escherichia coli. Arch Biochem Biophys. 1972 Jun;150(2):459–472. doi: 10.1016/0003-9861(72)90063-x. [DOI] [PubMed] [Google Scholar]
  23. Shipp W. S., Piotrowski M., Friedman A. E. Apparent cytochrome gene dose effects in F-lac and F-gal heterogenotes of Escherichia coli. Arch Biochem Biophys. 1972 Jun;150(2):473–481. doi: 10.1016/0003-9861(72)90064-1. [DOI] [PubMed] [Google Scholar]
  24. Van Wielink J. E., Oltmann L. F., Leeuwerik F. J., De Hollander J. A., Stouthamer A. H. A method for in situ characterization of b- and c-type cytochromes in Escherichia coli and in complex III from beef heart mitochondria by combined spectrum deconvolution and potentiometric analysis. Biochim Biophys Acta. 1982 Aug 20;681(2):177–190. doi: 10.1016/0005-2728(82)90021-4. [DOI] [PubMed] [Google Scholar]
  25. Wu T. T. A model for three-point analysis of random general transduction. Genetics. 1966 Aug;54(2):405–410. doi: 10.1093/genetics/54.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES