Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

PprA: a protein implicated in radioresistance of Deinococcus radiodurans stimulates catalase activity in Escherichia coli

Abstract

PprA: a pleiotropic protein promoting DNA repair, role in radiation resistance of Deinococcus radiodurans was demonstrated. In this study, the effect of radiation and oxidative stress on transgenic Escherichia coli expressing pprA has been studied. The pprA gene from D. radiodurans KR1 was cloned and expressed in E. coli. Transgenic E. coli cells expressing PprA showed twofold to threefold higher tolerance to hydrogen peroxide as compared to control. The 2.8-fold in vivo stimulation of catalase activity largely contributed by KatE was observed as compared to nonrecombinant control. Furthermore, the purified PprA could stimulate the E. coli catalase activity by 1.7-fold in solution. The effect of PprA on catalase activity observed both in vivo and in vitro was reverted to normal levels in the presence of PprA antibodies. The results suggest that enhanced oxidative stress tolerance in E. coli expressing PprA was due to the PprA stimulation of catalase activity, perhaps through the interaction of these proteins.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Anderson A, Nordan H, Cain R, Parrish G, Duggan D (1956) Studies on a radioresistant micrococcus. I. Isolation, morphology, cultural characteristics and resistance to gamma radiation. Food Technol 10:575–578

  2. Arrage AA, Phelps TJ, Benoit RE, White DC (1993) Survival of subsurface microorganisms exposed to UV radiation and hydrogen peroxide. Appl Environ Microbiol 59:3545–3550

  3. Battista JR (2000) Radiation resistance: the fragments that remain. Curr Biol 10:R204–R205

  4. Battista JR, Park M-J, McLemore AE (2001) Inactivation of two homologous of proteins presumed to be involved in the desiccation tolerance of plants sensitizes Deinococcus radiodurans R1 to desiccation. Cryobiology 42:133–139

  5. Beers Jr RF, Sizer IW (1951) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 159:133–140

  6. Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  7. Chou FI, Tan ST (1990) Manganese (II) induces cell division and increases in superoxide dismutase and catalase activities in an aging deinococcal culture. J Bacteriol 172:2029–2035

  8. Gao G, Tian B, Liu L, Sheng D, Shen B, Hua Y (2003) Expression of Deinococcus radiodurans PprI enhances the radioresistance of Escheichia coli. DNA Repair 2:1419–1427

  9. Halliwell B, Gutteridge MC (1999) Free radicals in Biology and Medicine. Oxford Univ Press, Oxford, pp 105–35

  10. Hua Y, Narumi I, Gao G, Tian B, Satoh K, Kitayama S, Shen B (2003) PprI: a general switch responsible for extreme radioresistance of Deinococcus radiodurans. Biochem Biophys Res Commun 306:354–360

  11. Khairnar NP, Misra HS, Apte SK (2003) Pyrroloquinoline–quinone synthesized in Escherichia coli by pyrroloquinoline–quinone synthase of Deinococcus radiodurans plays a role beyond mineral phosphate solubilization. Biochem Biophys Res Comm 312:303–308

  12. Kitayama S, Asaka S, Totsuka K (1983) DNA double-strand breakage and removal of cross-links in Deinococcus radiodurans. J Bacteriol 155:1200–1207

  13. Loewen PC, Switala J, Triggs-Raine BL (1985) Catalases HPI and HPII in Escherichia coli are induced independently. Arch Biochem Biophys 243:144–149

  14. Markillie LM, Varnum SM, Hradechy P, Wong KK (1999) Targeted mutagenesis by duplication insertion in the radioresistant bacterium Deinococcus radiodurans: radiation sensitivities of catalase (katA) and superoxide dismutase (sodA) mutants. J Bacteriol 181:666–669

  15. Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the auto-oxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474

  16. Minton KW (1994) DNA repair in the extremely radioresistant bacterium Deinococcus radiodurans. Mol Microbiol 13:9–15

  17. Misra HP, Fridovich I (1976) Superoxide dismutase and the oxygen enhancement of radiation lethality. Arch Biochem Biophys 176:577–581

  18. Misra HS, Khairnar NP, Barik A, Indirapriyadarsini K, Mohan H, Apte SK (2004) Pyrroloquinoline-quinone: a reactive oxygen species scavenger in bacteria. FEBS Lett 578:26–30

  19. Misra HS, Khairnar NP, Kota S, Shrivastava S, Joshi VP, Apte SK (2006) An Exonuclease I sensitive DNA repair pathways in Deinococcus radiodurans: a major determinant of radiation resistance. Mol Microbiol 59:1308–1316

  20. Mulder MA, Nair S, Abratt VR, Zappe H, Steyn LM (1999) Involvement of the N- and C-terminal domains of Mycobacterium tuberculosis KatG in the protection of mutant Escherichia coli against DNA-damaging agents. Microbiology 145:2011–2021

  21. Murray RGE (1986) Family II Deinococcaceae. In: PHA Sneath, NS Mair, ME Sharpe, J G Holt (eds) Bergey’s manual of systematic bacteriology, vol 2. The Williams and Wilkins, Baltimore, MD, pp 1035–104

  22. Narumi I, Satoh K, Cui S, Funayama T, Kitayama S, Watanabe H (2004) PprA: a novel protein from Deinococcus radiodurans that stimulates DNA ligation. Mol Microbiol 54:278–285

  23. Peak MJ, Peak JG, Foote CS, Krinsky NI (1984) Oxygen-independent direct deoxyribonucleic acid backbone breakage caused by Rose Bengal and visible light. J Photochem 25:309–315

  24. Sak BD, Eisenstark A, Touati D (1989) Exonuclease III and the catalase hydroperoxidase II in Escherichia coli are both regulated by the KatF gene product. Proc Natl Acad Sci USA 86:3271–3275

  25. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

  26. Smith MD, Masters CI, Moseley BEB (1992) Molecular Biology of radiation-resistant bacteria. In: RA Herbert and RJ Sharp (eds) Molecular biology and biotechnology of extremophiles. Chapman & Hall, New York, NY, pp 258–280

  27. Tian B, Wu Y, Sheng D, Zheng Z, Gao G, Hua Y (2004) Chemiluminescence assay for reactive oxygen species scavenging activities and inhibition on oxidative damage of DNA in Deinococcus radiodurans. Luminescence 19:78–84

  28. Wang P, Schellhorn HE (1995) Induction of resistance to hydrogen peroxide and radiation in Deinococcus radiodurans. Can J Microbiol 41:170–176

  29. Wayne LG, Diaz GA (1986) A double staining method for differentiating between two catalases of Mycobacterial catalase in polyacrylamide electrophoresis gels. Anal Biochem 157:89–92

  30. White O, Eisen JA, Heildelberg JF, Hickey EK, Peterson JD, Dodson RJ, Haft DH, Gwinn ML, Nelson WC, Richardson DL, Moffat KS, Qin H, Jiang L, Pamphile W, Crosby M, Shen M, Vamathevan JJ, Lam P, McDonald L, Utterback T, Zalewski C, Makarova KS, Aravind L, Daly MJ, Minton KW, Fleischmann RD, Ketchum KA, Nelson KE, Salzberg S, Smith HO, Venter JC, Fraser CM (1999) Genome sequence of the radioresistant bacterium Deinococcus radiodurans R1. Science 286:1571–1577

  31. Yun EJ, Lee YN (2000) Production of two different catalase–peroxidases by Deinococcus radiophilus. FEMS Microbiol Lett 184:155–159

Download references

Acknowledgements

Authors are thankful to Dr. Issay Narumi, JAERI, Japan, for a generous gift of Deinococcus radiodurans KR1 and PprA antibodies. Critical comments and suggestions from Dr. S. K. Apte and Dr. V. P. Joshi, during the preparation of the manuscript, are highly appreciated and deeply acknowledged.

Author information

Correspondence to Hari S. Misra.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kota, S., Misra, H.S. PprA: a protein implicated in radioresistance of Deinococcus radiodurans stimulates catalase activity in Escherichia coli . Appl Microbiol Biotechnol 72, 790–796 (2006). https://doi.org/10.1007/s00253-006-0340-7

Download citation

Keywords

  • Gamma Radiation
  • Catalase Activity
  • Oxidative Stress Response
  • Deinococcus Radiodurans
  • Oxidative Stress Tolerance