1887

Abstract

This work characterized the putative quinone oxidoreductase gene () from . The deduced amino acid sequence indicated that the 333 aa protein contains an NAD(P)H-binding motif. A Northern blot analysis revealed that 2·6 kb and 1·4 kb signals were detected by using a probe. Both the signals were enhanced under the presence of a redox-cycling agent, 9,10-phenanthrenequinone (PQ). It was also revealed that the expression of three genes, SA1988, SA1989 () and SA1990, was enhanced at the transcriptional level by PQ exposure. The results suggested that the 2·6 kb signal detected by the probe was in two co-transcripts, i.e. SA1990– and –SA1988 were transcribed. Besides, primer extension analyses confirmed the enhancement of and SA1990 transcripts. The GST (glutathione -transferase)-tagged QorA protein was expressed in and purified using a glutathione affinity column. In purification steps, a 36 kDa band co-purified with the GST–QorA, and it was detected even in the thrombin-cleaved fraction. N-terminal amino acid sequences for the 36 kDa protein revealed that it was an intact QorA. They showed that QorA formed a multimer under physiological conditions. The purified recombinant GST–QorA catalysed NADPH consumption in the presence of PQ as a substrate, but not NADH. To characterize the catalytic activity of QorA, superoxide anion that was generated through one-electron reduction of PQ and hydroquinone that was produced by two-electron reduction of PQ were measured. During reduction of PQ by GST–QorA, superoxide anion was generated, whereas a small amount of 9,10-dihydroxyphenanthrene (hydroquinone of PQ) was produced. These results suggest that the activity of QorA is similar to ζ-Crystallin, catalysing an NADPH-dependent one-electron reduction of quinone.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.25796-0
2003-02-01
2020-04-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/2/mic149389.html?itemId=/content/journal/micro/10.1099/mic.0.25796-0&mimeType=html&fmt=ahah

References

  1. Babiychuk E, Kushnir S, Belles-Boix E, Van Montagu M., Inze D. 1995; Arabidopsis thaliana NADPH oxidoreductase homologs confer tolerance of yeast toward the thiol-oxidizing drug diamide. J Biol Chem270:26224–26231
    [Google Scholar]
  2. Cadenas E. 1995; Antioxidant and prooxidant function of DT-diaphorase in quinone metabolism. Biochem Pharmacol49:127–140
    [Google Scholar]
  3. Clements M. O., Foster S. J. 1999; Stress resistance in Staphylococcus aureus. Trends Microbiol7:458–462
    [Google Scholar]
  4. Clements M. O, Watson S. P., Foster S. J. 1999; Characterization of the major superoxide dismutase of Staphylococcus aureus and its role in starvation survival, stress resistance, and pathogenicity. J Bacteriol181:3898–3903
    [Google Scholar]
  5. Edwards K. J, Barton J. D, Rossjohn J, Thorn J. M, Taylor G. L., Ollis D. L. 1996; Structural and sequence comparison of quinone oxidoreductase, ζ-Crystallin, and glucose and alcohol dehydrogenases. Arch Biochem Biophys328:173–183
    [Google Scholar]
  6. Ernster L. 1987; DT-diaphorase: a historical review. Chem Scr27A:1–13
    [Google Scholar]
  7. Friedrich T. 1998; The NADH : ubiquinone oxidoreductase (complex I) from Escherichia coli. Biochim Biophys Acta1364:134–136
    [Google Scholar]
  8. Hiramatsu K, Asada K, Suzuki E, Okonogi K., Yokota T. 1991; Molecular cloning and nucleotide sequence determination of the regulator region of mecA gene in methicillin-resistant Staphylococcus aureus (MRSA). FEBS Lett298:133–136
    [Google Scholar]
  9. Horsburgh M. J, Clements M. O, Crossley H, Ingham E., Foster S. J. 2001a; PerR controls oxidative stress resistance and iron storage proteins and is required for virulence in Staphylococcus aureus. Infect Immun69:3744–3754
    [Google Scholar]
  10. Horsburgh M. J, Ingham E., Foster S. J. 2001b; In Staphylococcus aureus , Fur is an interactive regulator with PerR, contributes to virulence, and is necessary for oxidative stress resistance through positive regulation of catalase and iron homeostasis. J Bacteriol183:468–475
    [Google Scholar]
  11. Huang Q.-L, Russell P, Stone S. H., Zigler J. S. Jr. 1987; Zeta-crystallin, a novel lens protein from guinea pig. Curr Eye Res6:725–732
    [Google Scholar]
  12. Iyanagi T. 1987; On the mechanisms of one- and two-electron transfer by flavin enzymes. Chem Scr27A:31–36
    [Google Scholar]
  13. Kumagai Y, Wakayama Y, Li S, Shinohara A, Iwamatsu A, Sun G., Shimojo N. 2000; ζ-Crystallin catalyzes the reductive activation of 2,4,6-trinitrotoluene to generate reactive oxygen species: a proposed mechanism for the induction of cataracts. FEBS Lett478:295–298
    [Google Scholar]
  14. Kunst F, Ogasawara N, Moszer I.. 148 other authors 1997; The complete genome sequence of the gram-positive bacterium Bacillus subtilis. Nature390:249–256
    [Google Scholar]
  15. Kuroda M, Ohta T, Uchiyama I.. 34 other authors 2001; Whole genome sequencing of methicillin-resistant Staphylococcus aureus. Lancet357:1225–1240
    [Google Scholar]
  16. Lilley P. E, Stamford N. P. J, Vasudevan S. G., Dixson N. E. 1993; The 92-min region of the Escherichia coli chromosome: location and cloning of the ubiA and alr genes. Gene129:9–16
    [Google Scholar]
  17. Mano J, Babiychuk E, Belles-Boix E, Hiratake J, Kimura A, Inze D, Kushnir S., Asada K. 2000; A novel NADPH : diamide oxidoreductase activity in Arabidopsis thaliana P1 ζ-Crystallin. Eur J Biochem267:3661–3671
    [Google Scholar]
  18. Ohta T, Noguchi S, Nakanishi M, Motoh Y, Hirata H, Kawamura M., Kagawa Y. 1991; The ‘lysine cluster’ in the N-terminal region of Na+/K(+)-ATPase alpha-subunit is not involved in ATPase activity. Biochim Biophys Acta1059:157–164
    [Google Scholar]
  19. Persson B, Zigler J. S. Jr, Jornvall H. 1994; A super-family of medium-chain dehydrogenases/reductases (MDR. Eur J Biochem226:15–22
    [Google Scholar]
  20. Rao P., Zigler J. S. Jr. 1992; Quinone-induced stimulation of hexose monophosphate shunt activity in the guinea pig lens: role of zeta-crystallin. Biochim Biophys Acta 1116;75–81
    [Google Scholar]
  21. Rao P. V, Krishna C. M., Zigler J. S. Jr. 1992; Identification and characterization of the enzymatic activity of ζ-Crystallin from guinea pig lens. J Biol Chem267:96–102
    [Google Scholar]
  22. Sanz R, Marín I, Ruiz-Santa-Quiteria J. A, Orden J. A, Cid D, Diez R. M, Silhadi K. S, Amils R., de la Fuente R. 2000; Catalase deficiency in Staphylococcus aureus subsp. anaerobius is associated with natural loss-of-function mutations within the structural gene. Microbiology146:465–475
    [Google Scholar]
  23. Thron J. M, Barton J. D, Dixon N. E, Ollis D. L., Edwards K. J. 1995; Crystal structure of Escherichia coli QOR quinone oxidoreductase complexed with NADPH. J Mol Biol249:785–799
    [Google Scholar]
  24. Valderas M. W., Hart M. 2001; Identification and characterization of a second superoxide dismutase gene ( sodM ) from Staphylococcus aureus. J Bacteriol183:3399–3407
    [Google Scholar]
  25. Yanisch-Perron C, Vieira J., Messing J. 1985; Improved M13phage cloning vectors and host strains: nucleotide sequence of the M13mp18 and pUC19 vectors. Gene33:103–119
    [Google Scholar]
  26. Yumoto I, Iwata H, Sawabe T, Ueno K, Ichise N, Matsuyama H, Okuyama H., Kawasaki K. 1999; Characterization of a facultatively psychrophilic bacterium, Vibrio rumoiensis sp. nov., that exhibits high catalase activity. Appl Environ Microbiol65:67–72
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.25796-0
Loading
/content/journal/micro/10.1099/mic.0.25796-0
Loading

Data & Media loading...

Most cited this month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error