1887

Microbiology Society logo

Volume 146, Issue 11

Control of periplasmic nitrate reductase gene expression () from in response to oxygen and carbon substrates Free

Abstract

The operon of encodes a periplasmic nitrate reductase (NAP), together with electron-transfer components and proteins required for the synthesis of a fully functional enzyme. Previously, it had been shown that high NAP activity was observed when was grown aerobically on highly reduced carbon sources such as butyrate or caproate, but not when cultured on more oxidized substrates such as succinate or malate. The enzyme is not present to any extent when the organism is grown anaerobically under denitrifying conditions, regardless of the carbon source. Transcriptional analyses of the operon have now identified two initiation sites which were differentially regulated in response to the carbon source, with expression being maximal when cells were grown aerobically with butyrate. Analysis of a mutant (M6) deregulated for NAP activity identified a single C→A transversion in a heptameric inverted-repeat sequence that partially overlapped the proximal promoter. Transcription analysis of this mutant revealed that expression of was completely derepressed under all growth conditions examined. Taken together, these findings indicate that transcription is negatively regulated during anaerobiosis, such that expression is restricted to aerobic growth, but only when the carbon source is highly reduced.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): MV+, reduced methyl viologen , NAP, periplasmic nitrate reductase , NAR, membrane-bound nitrate reductase , oxygen regulation , Paracoccus , Periplasmic nitrate reductase , repression and transcription
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-11-2977
2000-11-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/11/1462977a.html?itemId=/content/journal/micro/10.1099/00221287-146-11-2977&mimeType=html&fmt=ahah

References

  1. Bedzyk L., Wang T., Ye R. W. 1999; The periplasmic nitrate reductase in Pseudomonas sp. strain G-179 catalyses the first step of denitrification. J Bacteriol 181:2802–2806
     [Google Scholar]
  2. Bell L. C., Richardson D. J., Ferguson S. J. 1990; Periplasmic and membrane-bound respiratory nitrate reductases in Thiosphaera pantotropha: the periplasmic enzyme catalyses the first step in aerobic denitrification. FEBS Lett 265:85–87 [CrossRef]
     [Google Scholar]
  3. Bell L. C., Page M. D., Berks B. C., Richardson D. J., Ferguson S. J. 1993; Insertion of transposon Tn5 into a structural gene of the membrane-bound nitrate reductase of Thiosphaera pantotropha results in anaerobic overexpression of periplasmic nitrate reductase activity. J Gen Microbiol 139:3205–3214 [CrossRef]
     [Google Scholar]
  4. Berks B. C., Richardson D. J., Reilly A., Willis A. C., Ferguson S. J. 1995; The napEDABC cluster encoding the periplasmic nitrate reductase system of Thiosphaera pantotropha. Biochem J 309:983–992
     [Google Scholar]
  5. Blatny J. M., Brautaset T., Winther-Larsen H. C., Haugan K., Valla S. 1997; Construction and use of a versatile set of broad-host-range cloning and expression vectors based on the RK2 replicon. Appl Environ Microbiol 63:370–379
     [Google Scholar]
  6. Choy H., Adhya S. 1996; Negative control. In Escherichia coli and Salmonella: Cellular and Molecular Biology, 2nd edn. pp. 1287–1299Edited by Neidhardt F. C.others Washington, DC: American Society for Microbiology;
     [Google Scholar]
  7. Craske A. L., Ferguson S. J. 1986; The respiratory nitrate reductase from Paracoccus denitrificans: molecular characterisation and kinetic properties. Eur J Biochem 158:429–436 [CrossRef]
     [Google Scholar]
  8. Darwin A. J., Stewart V. 1995; Nitrate and nitrite regulation of the Fnr-dependent aeg-46.5 promoter of Escherichia coli K-12 is mediated by competition between homologous response regulators (NarL and NarP) for a common DNA-binding site. J Mol Biol 251:15–29 [CrossRef]
     [Google Scholar]
  9. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–563 [CrossRef]
     [Google Scholar]
  10. Figurski D. H., Helinski D. R. 1979; Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652 [CrossRef]
     [Google Scholar]
  11. Liu H. P., Takio S., Satoh T., Yamamoto I. 1999; Involvement in denitrification of the napKEFDABC genes encoding the periplasmic nitrate reductase system in the denitrifying phototrophic bacterium Rhodobacter sphaeroides f. sp. denitrificans. Biosci Biotechnol Biochem 63:530–536 [CrossRef]
     [Google Scholar]
  12. Potter L. C., Cole J. A. 1999; Essential roles for the products of the napABCD genes but not napFGH in periplasmic nitrate reduction by Escherichia coli K-12. Biochem J 344:69–76 [CrossRef]
     [Google Scholar]
  13. Potter L. C., Millington P., Griffiths L., Thomas G. H., Cole J. A. 1999; Competition between Escherichia coli strains expressing either a periplasmic or membrane-bound nitrate reductase: does NAP confer a selective advantage during nitrate-limited growth?. Biochem J 344:77–84 [CrossRef]
     [Google Scholar]
  14. Prentki P., Kritsch H. M. 1984; In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29:303–313 [CrossRef]
     [Google Scholar]
  15. Richardson D. J., Ferguson S. J. 1992; The influence of the carbon substrate on the activity of the periplasmic nitrate reductase in aerobically grown Thiosphaera pantotropha. Arch Microbiol 157:535–537
     [Google Scholar]
  16. Robertson L. A., Kuenen J. G. 1983; Thiosphaera pantotropha gen. nov. sp. nov., a facultatively anaerobic, facultatively autotrophic sulphur bacterium. J Gen Microbiol 129:2847–2855
     [Google Scholar]
  17. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467 [CrossRef]
     [Google Scholar]
  19. Sawers G. 1999; The aerobic/anaerobic interface. Curr Opin Microbiol 2:181–187 [CrossRef]
     [Google Scholar]
  20. Sawers G., Böck A. 1989; Novel transcriptional control of the pyruvate formate-lyase gene: upstream regulatory sequences and multiple promoters regulate anaerobic expression. J Bacteriol 171:2485–2498
     [Google Scholar]
  21. Sears H. J., Ferguson S. J., Richardson D. J., Spiro S. 1993; The identification of a periplasmic nitrate reductase in Paracoccus denitrificans. FEMS Microbiol Lett 113:107–112 [CrossRef]
     [Google Scholar]
  22. Sears H. J., Little P. J., Richardson D. J., Berks B. C., Spiro S., Ferguson S. J. 1997; Identification of an assimilatory nitrate reductase in mutants of Paracoccus denitrificans GB17 deficient in nitrate respiration. Arch Microbiol 167:61–66 [CrossRef]
     [Google Scholar]
  23. Siddiqui R. A., Warnecke-Eberz U., Hengsberger A., Schneider B., Kostka S., Friedrich B. 1993; Structure and function of periplasmic nitrate reductase in Alcaligenes eutrophus H16. J Bacteriol 175:5867–5876
     [Google Scholar]
  24. Spiro S. 1994; The FNR family of transcriptional regulators. Antonie Leeuwenhoek 66:23–36 [CrossRef]
     [Google Scholar]
  25. Van Spanning R. J. M., Wansell C. W., Harms N., Oltmann L. F., Stouthamer A. H. 1990; Mutagenesis of the gene encoding cytochrome c 550 of Paracoccus denitrificans and analysis of the resultant physiological effects. J Bacteriol 172:986–996
     [Google Scholar]
  26. Van Spanning R. J. M., Wansell C. W., Reijnders W. N. M., Harms N., Ras J., Otmann F., Stouthamer A. H. 1991; A method for introduction of unmarked mutations in the genome of Paracoccus denitrificans: construction of strains with multiple mutations in the genes encoding periplasmic cytochromes c 550, c 551i, and c 553i. J Bacteriol 173:6962–6970
     [Google Scholar]
  27. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268 [CrossRef]
     [Google Scholar]
  28. Wang H., Tseng C. P., Gunsalus R. P. 1999; The napF and narG nitrate reductase operons in Escherichia coli are differentially expressed in response to submicromolar concentrations of nitrate but not nitrite. J Bacteriol 181:5303–5308
     [Google Scholar]
  29. Yanish-Perron C., Vieira J., Messing J. 1985; Improved M13 cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-146-11-2977
Loading
Control of periplasmic nitrate reductase gene expression (napEDABC) from Paracoccus pantotrophus in response to oxygen and carbon substrates
Microbiology 146, 2977 (2000); https://doi.org/10.1099/00221287-146-11-2977
/content/journal/micro/10.1099/00221287-146-11-2977
/content/journal/micro/10.1099/00221287-146-11-2977
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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