1887

Abstract

aerobically respires with several terminal oxidases. The quinol oxidase (Cyo) encoded by is needed for efficient adaptation to low oxygen conditions and transcription is upregulated at low oxygen. This study sought to determine how transcription of the operon is regulated. The 5′ sequence upstream of was analysed and revealed putative binding sites for ActR of the ActSR two-component regulatory system. The expression of was decreased in an mutant regardless of the oxygen condition. As ActSR is known to be important for growth under low pH in another rhizobial species, the effect of growth medium pH on expression was tested. As the pH of the media was incrementally decreased, expression gradually increased in the WT, eventually reaching ∼10-fold higher levels at low pH (4.8) compared with neutral pH (7.0) conditions. This upregulation of under decreasing pH conditions was eliminated in the mutant. Both the and mutants had severe growth defects at low pH (4.8). Lastly, the and mutants had severe growth defects when grown in media treated with an iron chelator. Under these conditions, was upregulated in the WT, whereas was not induced in the mutant. Altogether, the results indicated expression is largely dependent on the ActSR two-component system. This study also demonstrated additional physiological roles for Cyo in CFN42, in which it is the preferred oxidase for growth under acidic and low iron conditions.

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2015-09-01
2019-10-23
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References

  1. Alexeyev M. F. . ( 1995;). Three kanamycin resistance gene cassettes with different polylinkers. Biotechniques 18: 52–56 [PubMed].
    [Google Scholar]
  2. Bauer E. , Kaspar T. , Fischer H. M. , Hennecke H. . ( 1998;). Expression of the fixRnifA operon in Bradyrhizobium japonicum depends on a new response regulator. Reg R. J Bacteriol 180: 3853–3863 [PubMed].
    [Google Scholar]
  3. Bobik C. , Meilhoc E. , Batut J. . ( 2006;). FixJ: a major regulator of the oxygen limitation response and late symbiotic functions of Sinorhizobium meliloti . J Bacteriol 188: 4890–4902 [CrossRef] [PubMed].
    [Google Scholar]
  4. Bueno E. , Mesa S. , Bedmar E. J. , Richardson D. J. , Delgado M. J. . ( 2012;). Bacterial adaptation of respiration from oxic to microoxic and anoxic conditions: redox control. Antioxid Redox Signal 16: 819–852 [CrossRef] [PubMed].
    [Google Scholar]
  5. Chao T. C. , Buhrmester J. , Hansmeier N. , Pühler A. , Weidner S. . ( 2005;). Role of the regulatory gene rirA in the transcriptional response of Sinorhizobium meliloti to iron limitation. Appl Environ Microbiol 71: 5969–5982 [CrossRef] [PubMed].
    [Google Scholar]
  6. Crofts A. R. , Hong S. , Ugulava N. , Barquera B. , Gennis R. , Guergova-Kuras M. , Berry E. A. . ( 1999;). Pathways for proton release during ubihydroquinone oxidation by the bc 1 complex. Proc Natl Acad Sci U S A 96: 10021–10026 [CrossRef] [PubMed].
    [Google Scholar]
  7. Dombrecht B. , Vanderleyden J. , Michiels J. . ( 2001;). Stable RK2-derived cloning vectors for the analysis of gene expression and gene function in gram-negative bacteria. Mol Plant Microbe Interact 14: 426–430 [CrossRef] [PubMed].
    [Google Scholar]
  8. Elsen S. , Swem L. R. , Swem D. L. , Bauer C. E. . ( 2004;). RegB/RegA, a highly conserved redox-responding global two-component regulatory system. Microbiol Mol Biol Rev 68: 263–279 [CrossRef] [PubMed].
    [Google Scholar]
  9. Emmerich R. , Hennecke H. , Fischer H. M. . ( 2000a;). Evidence for a functional similarity between the two-component regulatory systems RegSR, ActSR, and RegBA (PrrBA) in α-Proteobacteria. Arch Microbiol 174: 307–313 [CrossRef] [PubMed].
    [Google Scholar]
  10. Emmerich R. , Strehler P. , Hennecke H. , Fischer H. M. . ( 2000b;). An imperfect inverted repeat is critical for DNA binding of the response regulator RegR of Bradyrhizobium japonicum . Nucleic Acids Res 28: 4166–4171 [CrossRef] [PubMed].
    [Google Scholar]
  11. Finan T. M. , Kunkel B. , De Vos G. F. , Signer E. R. . ( 1986;). Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and thiamine synthesis genes. J Bacteriol 167: 66–72 [PubMed].
    [Google Scholar]
  12. Finan T. M. , Weidner S. , Wong K. , Buhrmester J. , Chain P. , Vorhölter F. J. , Hernandez-Lucas I. , Becker A. , Cowie A. , other authors . ( 2001;). The complete sequence of the 1,683-kb pSymB megaplasmid from the N2-fixing endosymbiont Sinorhizobium meliloti . Proc Natl Acad Sci U S A 98: 9889–9894 [CrossRef] [PubMed].
    [Google Scholar]
  13. Gao T. , O'Brian M. R. . ( 2005;). Iron-dependent cytochrome c 1 expression is mediated by the status of heme in Bradyrhizobium japonicum . J Bacteriol 187: 5084–5089 [CrossRef] [PubMed].
    [Google Scholar]
  14. García-Horsman J. A. , Barquera B. , Rumbley J. , Ma J. , Gennis R. B. . ( 1994;). The superfamily of heme-copper respiratory oxidases. J Bacteriol 176: 5587–5600 [PubMed].
    [Google Scholar]
  15. Glazebrook J. , Walker G. C. . ( 1991;). Genetic techniques in Rhizobium meliloti . Methods Enzymol 204: 398–418 [CrossRef] [PubMed].
    [Google Scholar]
  16. González V. , Santamaría R. I. , Bustos P. , Hernández-González I. , Medrano-Soto A. , Moreno-Hagelsieb G. , Janga S. C. , Ramírez M. A. , Jiménez-Jacinto V. , other authors . ( 2006;). The partitioned Rhizobium etli genome: genetic and metabolic redundancy in seven interacting replicons. Proc Natl Acad Sci U S A 103: 3834–3839 [CrossRef] [PubMed].
    [Google Scholar]
  17. Guergova-Kuras M. , Kuras R. , Ugulava N. , Hadad I. , Crofts A. R. . ( 2000;). Specific mutagenesis of the Rieske iron-sulfur protein in Rhodobacter sphaeroides shows that both the thermodynamic gradient and the pK of the oxidized form determine the rate of quinol oxidation by the bc 1 complex. Biochemistry 39: 7436–7444 [CrossRef] [PubMed].
    [Google Scholar]
  18. Hanahan D. . ( 1983;). Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166: 557–580 [CrossRef] [PubMed].
    [Google Scholar]
  19. Hellweg C. , Pühler A. , Weidner S. . ( 2009;). The time course of the transcriptomic response of Sinorhizobium meliloti 1021 following a shift to acidic pH. BMC Microbiol 9: 37 [CrossRef] [PubMed].
    [Google Scholar]
  20. Hoang T. T. , Karkhoff-Schweizer R. R. , Kutchma A. J. , Schweizer H. P. . ( 1998;). A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 212: 77–86 [CrossRef] [PubMed].
    [Google Scholar]
  21. Kaneko T. , Nakamura Y. , Sato S. , Minamisawa K. , Uchiumi T. , Sasamoto S. , Watanabe A. , Idesawa K. , Iriguchi M. , other authors . ( 2002;). Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110. DNA Res 9: 189–197 [CrossRef] [PubMed].
    [Google Scholar]
  22. Kawakami T. , Kuroki M. , Ishii M. , Igarashi Y. , Arai H. . ( 2010;). Differential expression of multiple terminal oxidases for aerobic respiration in Pseudomonas aeruginosa . Environ Microbiol 12: 1399–1412 [PubMed].
    [Google Scholar]
  23. Kiley P. J. , Beinert H. . ( 1998;). Oxygen sensing by the global regulator, FNR: the role of the iron-sulfur cluster. FEMS Microbiol Rev 22: 341–352 [CrossRef] [PubMed].
    [Google Scholar]
  24. Körner H. , Sofia H. J. , Zumft W. G. . ( 2003;). Phylogeny of the bacterial superfamily of Crp-Fnr transcription regulators: exploiting the metabolic spectrum by controlling alternative gene programs. FEMS Microbiol Rev 27: 559–592 [CrossRef] [PubMed].
    [Google Scholar]
  25. Lhee S. , Kolling D. R. , Nair S. K. , Dikanov S. A. , Crofts A. R. . ( 2010;). Modifications of protein environment of the [2Fe-2S] cluster of the bc 1 complex: effects on the biophysical properties of the Rieske iron-sulfur protein and on the kinetics of the complex. J Biol Chem 285: 9233–9248 [CrossRef] [PubMed].
    [Google Scholar]
  26. Lindemann A. , Moser A. , Pessi G. , Hauser F. , Friberg M. , Hennecke H. , Fischer H. M. . ( 2007;). New target genes controlled by the Bradyrhizobium japonicum two-component regulatory system RegSR. J Bacteriol 189: 8928–8943 [CrossRef] [PubMed].
    [Google Scholar]
  27. Lunak Z. R. , Noel K. D. . ( 2015;). A quinol oxidase, encoded by cyoABCD, is utilized to adapt to lower O2 concentrations in Rhizobium etli CFN42. Microbiology 161: 203–212 [PubMed].[CrossRef]
    [Google Scholar]
  28. Morris R. L. , Schmidt T. M. . ( 2013;). Shallow breathing: bacterial life at low O2 . Nat Rev Microbiol 11: 205–212 [CrossRef] [PubMed].
    [Google Scholar]
  29. Noel K. D. , Sanchez A. , Fernandez L. , Leemans J. , Cevallos M. A. . ( 1984;). Rhizobium phaseoli symbiotic mutants with transposon Tn5 insertions. J Bacteriol 158: 148–155 [PubMed].
    [Google Scholar]
  30. O'Hara G. W. , Goss T. J. , Dilworth M. J. , Glenn A. R. . ( 1989;). Maintenance of intracellular pH and acid tolerance in Rhizobium meliloti . Appl Environ Microbiol 55: 1870–1876 [PubMed].
    [Google Scholar]
  31. Ojeda K. J. , Box J. M. , Noel K. D. . ( 2010;). Genetic basis for Rhizobium etli CE3 O-antigen O-methylated residues that vary according to growth conditions. J Bacteriol 192: 679–690 [CrossRef] [PubMed].
    [Google Scholar]
  32. Rudolph G. , Hennecke H. , Fischer H. M. . ( 2006;). Beyond the Fur paradigm: iron-controlled gene expression in rhizobia. FEMS Microbiol Rev 30: 631–648 [CrossRef] [PubMed].
    [Google Scholar]
  33. 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]
  34. Spaink H. P. , Okker R. J. H. , Wijffelman C. A. , Pees E. , Lugtenberg B. J. J. . ( 1987;). Promoters in the nodulation region of the Rhizobium leguminosarum Sym plasmid pRL1JI. Plant Mol Biol 9: 27–39 [CrossRef] [PubMed].
    [Google Scholar]
  35. Surpin M. A. , Maier R. J. . ( 1998;). Roles of the Bradyrhizobium japonicum terminal oxidase complexes in microaerobic H2-dependent growth. Biochim Biophys Acta 1364: 37–45 [CrossRef] [PubMed].
    [Google Scholar]
  36. Thompson J. D. , Gibson T. J. , Higgins D. G. . ( 2002;). Multiple sequence alignment using Clustal W and Clustal X. Curr Protoc Bioinformatics Chapter 2: 3 [CrossRef] [PubMed].
    [Google Scholar]
  37. Tiwari R. P. , Reeve W. G. , Dilworth M. J. , Glenn A. R. . ( 1996;). Acid tolerance in Rhizobium meliloti strain WSM419 involves a two-component sensor-regulator system. Microbiology 142: 1693–1704 [CrossRef] [PubMed].
    [Google Scholar]
  38. Torres M. J. , Argandoña M. , Vargas C. , Bedmar E. J. , Fischer H. M. , Mesa S. , Delgado M. J. . ( 2014;). The global response regulator RegR controls expression of denitrification genes in Bradyrhizobium japonicum . PLoS One 9: e99011 [PubMed].[CrossRef]
    [Google Scholar]
  39. Trzebiatowski J. R. , Ragatz D. M. , de Bruijn F. J. . ( 2001;). Isolation and regulation of Sinorhizobium meliloti 1021 loci induced by oxygen limitation. Appl Environ Microbiol 67: 3728–3731 [CrossRef] [PubMed].
    [Google Scholar]
  40. Wu J. , Bauer C. E. . ( 2008;). RegB/RegA, a global redox-responding two-component system. Adv Exp Med Biol 631: 131–148 [CrossRef] [PubMed].
    [Google Scholar]
  41. Wu J. , Bauer C. E. . ( 2010;). RegB kinase activity is controlled in part by monitoring the ratio of oxidized to reduced ubiquinones in the ubiquinone pool. MBio 1: e00272-10 [CrossRef] [PubMed].
    [Google Scholar]
  42. Young J. P. , Crossman L. C. , Johnston A. W. , Thomson N. R. , Ghazoui Z. F. , Hull K. H. , Wexler M. , Curson A. R. , Todd J. D. , other authors . ( 2006;). The genome of Rhizobium leguminosarum has recognizable core and accessory components. Genome Biol 7: R34 [CrossRef] [PubMed].
    [Google Scholar]
  43. Zhou F. , Yin Y. , Su T. , Yu L. , Yu C. A. . ( 2012;). Oxygen dependent electron transfer in the cytochrome bc 1 complex. Biochim Biophys Acta 1817: 2103–2109 [CrossRef] [PubMed].
    [Google Scholar]
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