1887

Abstract

The ActSR two-component regulatory system is a member of a homologous group of global redox-responsive regulatory systems that adjust the expression of energy-consuming and energy-supplying metabolic pathways in order to maintain cellular redox balance. In this study, the transcriptional organization of the locus was determined and the effect of system inactivation on stress resistance was investigated. It was found that is transcribed as a monocistronic mRNA and is transcribed along with as a bicistronic mRNA, while is also transcribed as a monocistronic message. Each message is initiated from a separate promoter. Inactivation of resulted in decreased resistance to membrane stress (sodium dodecyl sulfate), acid stress (pH 5.5), iron starvation (bipyridyl) and iron excess (FeCl), and antibiotic stress (tetracycline and ciprofloxacin). Resistance to oxidative stress in the form of organic peroxide (cumene hydroperoxide) increased, while resistance to inorganic peroxide (HO) decreased. An insertion mutant displayed reduced catalase activity, even though transcription of and remained unchanged. Complementation of the inactivation mutant with plasmid-encoded or overexpression of , encoding ferrochelatase, restored wild-type catalase activity and HO resistance levels. Gel mobility shift and promoter fusion results indicated that ActR is a positive regulator of that binds directly to the promoter region. Thus, inactivation of the ActSR system affects resistance to multiple stresses, including reduced resistance to HO resulting from a reduction in catalase activity due to reduced expression of .

Keyword(s): ActR , ActS , heme , hydrogen peroxide and stress
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/content/journal/micro/10.1099/mic.0.000838
2019-10-01
2019-10-15
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References

  1. Dubbs JM, Tabita FR. Regulators of nonsulfur purple phototrophic bacteria and the interactive control of CO2 assimilation, nitrogen fixation, hydrogen metabolism and energy generation. FEMS Microbiol Rev 2004;28:353–376 [CrossRef]
    [Google Scholar]
  2. Emmerich R, Hennecke H, Fischer HM. Evidence for a functional similarity between the two-component regulatory systems RegSR, ActSR, and RegBA (PrrBA) in alpha-Proteobacteria. Arch Microbiol 2000;174:307–313 [CrossRef]
    [Google Scholar]
  3. Comolli JC, Donohue TJ. Pseudomonas aeruginosa RoxR, a response regulator related to Rhodobacter sphaeroides PrrA, activates expression of the cyanide-insensitive terminal oxidase. Mol Microbiol 2002;45:755–768 [CrossRef]
    [Google Scholar]
  4. Elsen S, Swem LR, Swem DL, Bauer CE. RegB/RegA, a highly conserved redox-responding global two-component regulatory system. Microbiol Mol Biol Rev 2004;68:263–279 [CrossRef]
    [Google Scholar]
  5. Eraso JM, Kaplan S. Oxygen-insensitive synthesis of the photosynthetic membranes of Rhodobacter sphaeroides: a mutant histidine kinase. J Bacteriol 1995;177:2695–2706 [CrossRef]
    [Google Scholar]
  6. Mosley CS, Suzuki JY, Bauer CE. Identification and molecular genetic characterization of a sensor kinase responsible for coordinately regulating light harvesting and reaction center gene expression in response to anaerobiosis. J Bacteriol 1994;176:7566–7573 [CrossRef]
    [Google Scholar]
  7. Eraso JM, Kaplan S. prrA, a putative response regulator involved in oxygen regulation of photosynthesis gene expression in Rhodobacter sphaeroides. J Bacteriol 1994;176:32–43 [CrossRef]
    [Google Scholar]
  8. Sganga MW, Bauer CE. Regulatory factors controlling photosynthetic reaction center and light-harvesting gene expression in Rhodobacter capsulatus. Cell 1992;68:945–954 [CrossRef]
    [Google Scholar]
  9. Eraso JM, Roh JH, Zeng X, Callister SJ, Lipton MS et al. Role of the global transcriptional regulator PrrA in Rhodobacter sphaeroides 2.4.1: combined transcriptome and proteome analysis. J Bacteriol 2008;190:4831–4848 [CrossRef]
    [Google Scholar]
  10. Bird TH, Du S, Bauer CE. Autophosphorylation, phosphotransfer, and DNA-binding properties of the RegB/RegA two-component regulatory system in Rhodobacter capsulatus. J Biol Chem 1999;274:16343–16348 [CrossRef]
    [Google Scholar]
  11. Seok J-S, Kaplan S, Oh J-I. Interacting specificity of a histidine kinase and its cognate response regulator: the PrrBA system of Rhodobacter sphaeroides. Microbiology 2006;152:2479–2490 [CrossRef]
    [Google Scholar]
  12. Oh J-I, Ko I-J, Kaplan S, JI O. Reconstitution of the Rhodobacter sphaeroides cbb3-PrrBA signal transduction pathway in vitro. Biochemistry 2004;43:7915–7923 [CrossRef]
    [Google Scholar]
  13. Potter CA, Ward A, Laguri C, Williamson MP, Henderson PJF et al. Expression, purification and characterisation of full-length histidine protein kinase RegB from Rhodobacter sphaeroides. J Mol Biol 2002;320:201–213 [CrossRef]
    [Google Scholar]
  14. Wu J, Bauer CE. Regb kinase activity is controlled in part by monitoring the ratio of oxidized to reduced ubiquinones in the ubiquinone pool. MBio 2010;1:e00272-10 [CrossRef]
    [Google Scholar]
  15. Wu J, Cheng Z, Reddie K, Carroll K, Hammad LA et al. Regb kinase activity is repressed by oxidative formation of cysteine sulfenic acid. J Biol Chem 2013;288:4755–4762 [CrossRef]
    [Google Scholar]
  16. Kim Y-J, Ko I-J, Lee J-M, Kang H-Y, Kim YM et al. Dominant role of the cbb3 oxidase in regulation of photosynthesis gene expression through the PrrBA system in Rhodobacter sphaeroides 2.4.1. J Bacteriol 2007;189:5617–5625 [CrossRef]
    [Google Scholar]
  17. Laguri C, Stenzel RA, Donohue TJ, Phillips-Jones MK, Williamson MP. Activation of the global gene regulator PrrA (RegA) from Rhodobacter sphaeroides. Biochemistry 2006;45:7872–7881 [CrossRef]
    [Google Scholar]
  18. Schindel HS, Bauer CE. The RegA regulon exhibits variability in response to altered growth conditions and differs markedly between Rhodobacter species. Microb Genom 2016;2:e000081 [CrossRef]
    [Google Scholar]
  19. Imam S, Noguera DR, Donohue TJ. Global analysis of photosynthesis transcriptional regulatory networks. PLoS Genet 2014;10:e1004837 [CrossRef]
    [Google Scholar]
  20. Lindemann A, Moser A, Pessi G, Hauser F, Friberg M et al. New target genes controlled by the Bradyrhizobium japonicum two-component regulatory system RegSR. J Bacteriol 2007;189:8928–8943 [CrossRef]
    [Google Scholar]
  21. Torres MJ, Argandoña M, Vargas C, Bedmar EJ, Fischer H-M et al. The global response regulator RegR controls expression of denitrification genes in Bradyrhizobium japonicum. PLoS One 2014;9:e99011 [CrossRef]
    [Google Scholar]
  22. Dangel AW, Luther A, Tabita FR. Amino acid residues of RegA important for interactions with the CbbR-DNA complex of Rhodobacter sphaeroides. J Bacteriol 2014;196:3179–3190 [CrossRef]
    [Google Scholar]
  23. Gonzalez-Mula A, Lachat J, Mathias L, Naquin D, Lamouche F et al. The biotroph Agrobacterium tumefaciens thrives in tumors by exploiting a wide spectrum of plant host metabolites. New Phytol 2018
    [Google Scholar]
  24. Yuan Z-C, Liu P, Saenkham P, Kerr K, Nester EW. Transcriptome profiling and functional analysis of Agrobacterium tumefaciens reveals a general conserved response to acidic conditions (pH 5.5) and a complex acid-mediated signaling involved in Agrobacterium-plant interactions. J Bacteriol 2008;190:494–507 [CrossRef]
    [Google Scholar]
  25. Ngok-Ngam P, Ruangkiattikul N, Mahavihakanont A, Virgem SS, Sukchawalit R et al. Roles of Agrobacterium tumefaciens RirA in iron regulation, oxidative stress response, and virulence. J Bacteriol 2009;191:2083–2090 [CrossRef]
    [Google Scholar]
  26. Kitphati W, Ngok-Ngam P, Suwanmaneerat S, Sukchawalit R, Mongkolsuk S. Agrobacterium tumefaciens fur has important physiological roles in iron and manganese homeostasis, the oxidative stress response, and full virulence. Appl Environ Microbiol 2007;73:4760–4768 [CrossRef]
    [Google Scholar]
  27. Imlay JA. Pathways of oxidative damage. Annu Rev Microbiol 2003;57:395–418 [CrossRef]
    [Google Scholar]
  28. Jalloul A, Montillet JL, Assigbetsé K, Agnel JP, Delannoy E et al. Lipid peroxidation in cotton: Xanthomonas interactions and the role of lipoxygenases during the hypersensitive reaction. Plant J 2002;32:1–12 [CrossRef]
    [Google Scholar]
  29. Bolwell GP. Role of active oxygen species and NO in plant defence responses. Curr Opin Plant Biol 1999;2:287–294 [CrossRef]
    [Google Scholar]
  30. Gohlke J, Deeken R. Plant responses to Agrobacterium tumefaciens and crown gall development. Front Plant Sci 2014;5:155 [CrossRef]
    [Google Scholar]
  31. Baek S-H, Hartsock A, Shapleigh JP. Agrobacterium tumefaciens C58 uses ActR and FnrN to control nirK and nor expression. J Bacteriol 2008;190:78–86 [CrossRef]
    [Google Scholar]
  32. Davies BW, Walker GC. Identification of novel Sinorhizobium meliloti mutants compromised for oxidative stress protection and symbiosis. J Bacteriol 2007;189:2110–2113 [CrossRef]
    [Google Scholar]
  33. Tang G, Wang S, Lu D, Huang L, Li N et al. Two-component regulatory system ActS/ActR is required for Sinorhizobium meliloti adaptation to oxidative stress. Microbiol Res 2017;198:1–7 [CrossRef]
    [Google Scholar]
  34. Luo ZQ, Clemente TE, Farrand SK. Construction of a derivative of Agrobacterium tumefaciens C58 that does not mutate to tetracycline resistance. Mol Plant Microbe Interact 2001;14:98–103 [CrossRef]
    [Google Scholar]
  35. Metcalf WW, Jiang W, Daniels LL, Kim SK, Haldimann A et al. Conditionally replicative and conjugative plasmids carrying lacZ alpha for cloning, mutagenesis, and allele replacement in bacteria. Plasmid 1996;35:1–13 [CrossRef]
    [Google Scholar]
  36. Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 1985;33:103–119 [CrossRef]
    [Google Scholar]
  37. Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP. 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 1998;212:77–86 [CrossRef]
    [Google Scholar]
  38. Marx CJ, Lidstrom ME. Broad-Host-Range cre-lox system for antibiotic marker recycling in gram-negative bacteria. Biotechniques 2002;33:1062–1067 [CrossRef]
    [Google Scholar]
  39. Kovach ME, Elzer PH, Steven Hill D, Robertson GT, Farris MA et al. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 1995;166:175–176 [CrossRef]
    [Google Scholar]
  40. Spaink HP, Okker RJ, Wijffelman CA, Pees E, Lugtenberg BJ. Promoters in the nodulation region of the Rhizobium leguminosarum Sym plasmid pRL1JI. Plant Mol Biol 1987;9:27–39 [CrossRef]
    [Google Scholar]
  41. Hwang I, Cook DM, Farrand SK. A new regulatory element modulates homoserine lactone-mediated autoinduction of Ti plasmid conjugal transfer. J Bacteriol 1995;177:449–458 [CrossRef]
    [Google Scholar]
  42. Alexeyev MF, Shokolenko IN, Croughan TP. Improved antibiotic-resistance gene cassettes and omega elements for Escherichia coli vector construction and in vitro deletion/insertion mutagenesis. Gene 1995;160:63–67 [CrossRef]
    [Google Scholar]
  43. Bhubhanil S, Niamyim P, Sukchawalit R, Mongkolsuk S. Cysteine desulphurase-encoding gene sufS2 is required for the repressor function of RirA and oxidative resistance in Agrobacterium tumefaciens. Microbiology 2014;160:79–90 [CrossRef]
    [Google Scholar]
  44. Liu P, Wood D, Nester EW. Phosphoenolpyruvate carboxykinase is an acid-induced, chromosomally encoded virulence factor in Agrobacterium tumefaciens. J Bacteriol 2005;187:6039–6045 [CrossRef]
    [Google Scholar]
  45. Solovyev VS.Automatic annotation of microbial genomes and metagenomic sequences In Metagenomics and its Applications in Agriculture, Biomedicine and Environmental Studies (Ed RW Li), ISBN: 978-1-61668-682-6. Nova Science Publishers; 2011; pp61–78
    [Google Scholar]
  46. Mongkolsuk S, Loprasert S, Vattanaviboon P, Chanvanichayachai C, Chamnongpol S et al. Heterologous growth phase- and temperature-dependent expression and H2O2 toxicity protection of a superoxide-inducible monofunctional catalase gene from Xanthomonas oryzae pv. oryzae. J Bacteriol 1996;178:3578–3584 [CrossRef]
    [Google Scholar]
  47. Sambrook J, Russell DW. Molecular Cloning : a Laboratory Manual, Third edition. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 2001
    [Google Scholar]
  48. Beers RF, Sizer IW. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 1952;195:133–140
    [Google Scholar]
  49. Ochsner UA, Hassett DJ, Vasil ML. Genetic and physiological characterization of Ohr, encoding a protein involved in organic hydroperoxide resistance in Pseudomonas aeruginosa. J Bacteriol 2001;183:773–778 [CrossRef]
    [Google Scholar]
  50. Dokpikul T, Chaoprasid P, Saninjuk K, Sirirakphaisarn S, Johnrod J et al. Regulation of the cobalt/nickel efflux operon dmeRF in Agrobacterium tumefaciens and a Link between the Iron-Sensing Regulator RirA and Cobalt/Nickel resistance. Appl Environ Microbiol 2016;82:4732–4742 [CrossRef]
    [Google Scholar]
  51. Gautheret D, Lambert A. Direct RNA motif definition and identification from multiple sequence alignments using secondary structure profiles. J Mol Biol 2001;313:1003–1011 [CrossRef]
    [Google Scholar]
  52. Macke TJ et al. RNAMotif, an RNA secondary structure definition and search algorithm. Nucleic Acids Res 2001;29:4724–4735 [CrossRef]
    [Google Scholar]
  53. Bauer E, Kaspar T, Fischer HM, Hennecke H. Expression of the fixR-nifA operon in Bradyrhizobium japonicum depends on a new response regulator, RegR. J Bacteriol 1998;180:3853–3863
    [Google Scholar]
  54. Ewann F, Jackson M, Pethe K, Cooper A, Mielcarek N et al. Transient requirement of the PrrA-PrrB two-component system for early intracellular multiplication of Mycobacterium tuberculosis. Infect Immun 2002;70:2256–2263 [CrossRef]
    [Google Scholar]
  55. Ranson-Olson B, Zeilstra-Ryalls JH. Regulation of the Rhodobacter sphaeroides 2.4.1 hemA gene by PrrA and FnrL. J Bacteriol 2008;190:6769–6778 [CrossRef]
    [Google Scholar]
  56. Smart JL, Willett JW, Bauer CE. Regulation of hem gene expression in Rhodobacter capsulatus by redox and photosystem regulators RegA, CrtJ, FnrL, and AerR. J Mol Biol 2004;342:1171–1186 [CrossRef]
    [Google Scholar]
  57. Chelikani P, Fita I, Loewen PC. Diversity of structures and properties among catalases. Cell Mol Life Sci 2004;61:192–208 [CrossRef]
    [Google Scholar]
  58. Dailey HA, Dailey TA, Gerdes S, Jahn D, Jahn M et al. Prokaryotic heme biosynthesis: multiple pathways to a common essential product. Microbiol Mol Biol Rev 2017;81: [CrossRef]
    [Google Scholar]
  59. Eraso JM, Kaplan S. Half-Site DNA sequence and spacing length contributions to PrrA binding to PrrA site 2 of RSP3361 in Rhodobacter sphaeroides 2.4.1. J Bacteriol 2009;191:4353–4364 [CrossRef]
    [Google Scholar]
  60. Mao L et al. Combining microarray and genomic data to predict DNA binding motifs. Microbiology 2005;151:3197–3213 [CrossRef]
    [Google Scholar]
  61. Dailey HA, Dailey TA, Gerdes S, Jahn D, Jahn M et al. Prokaryotic heme biosynthesis: multiple pathways to a common essential product. Microbiol Mol Biol Rev 2017;81: [CrossRef]
    [Google Scholar]
  62. Tiwari RP, Reeve WG, Dilworth MJ, Glenn AR. Acid tolerance in Rhizobium meliloti strain WSM419 involves a two-component sensor-regulator system. Microbiology 1996;142:1693–1704 [CrossRef]
    [Google Scholar]
  63. Du S, Kouadio JL, Bauer CE. Regulated expression of a highly conserved regulatory gene cluster is necessary for controlling photosynthesis gene expression in response to anaerobiosis in Rhodobacter capsulatus. J Bacteriol 1999;181:4334–4341
    [Google Scholar]
  64. Fernández-Piñar R, Ramos JL, Rodríguez-Herva JJ, Espinosa-Urgel M. A two-component regulatory system integrates redox state and population density sensing in Pseudomonas putida. J Bacteriol 2008;190:7666–7674 [CrossRef]
    [Google Scholar]
  65. Frank DW, Storey DG, Hindahl MS, Iglewski BH. Differential regulation by iron of regA and toxA transcript accumulation in Pseudomonas aeruginosa. J Bacteriol 1989;171:5304–5313 [CrossRef]
    [Google Scholar]
  66. Storey DG, Raivio TL, Frank DW, Wick MJ, Kaye S et al. Effect of regB on expression from the P1 and P2 promoters of the Pseudomonas aeruginosa regAB operon. J Bacteriol 1991;173:6088–6094 [CrossRef]
    [Google Scholar]
  67. Dubbs JM, Mongkolsuk S. Peroxide-sensing transcriptional regulators in bacteria. J Bacteriol 2012;194:5495–5503 [CrossRef]
    [Google Scholar]
  68. Charoenlap N, Eiamphungporn W, Chauvatcharin N, Utamapongchai S, Vattanaviboon P et al. OxyR mediated compensatory expression between ahpC and katA and the significance of ahpC in protection from hydrogen peroxide in Xanthomonas campestris. FEMS Microbiol Lett 2005;249:73–78 [CrossRef]
    [Google Scholar]
  69. Lindemann A, Koch M, Pessi G, Müller AJ, Balsiger S et al. Host-specific symbiotic requirement of BdeAB, a RegR-controlled RND-type efflux system in Bradyrhizobium japonicum. FEMS Microbiol Lett 2010;312:184–191 [CrossRef]
    [Google Scholar]
  70. Reeve WG, Tiwari RP, Dilworth MJ, Glenn AR. A helicase gene (helO) in Rhizobium meliloti WSM419. FEMS Microbiol Lett 1997;153:43–49 [CrossRef]
    [Google Scholar]
  71. Granato LM, Picchi SC, Andrade MdeO, Takita MA, de Souza AA et al. The ATP-dependent RNA helicase HrpB plays an important role in motility and biofilm formation in Xanthomonas citri subsp. citri. BMC Microbiol 2016;16:55 [CrossRef]
    [Google Scholar]
  72. Mancini S, Imlay JA. The induction of two biosynthetic enzymes helps Escherichia coli sustain heme synthesis and activate catalase during hydrogen peroxide stress. Mol Microbiol 2015;96:744–763 [CrossRef]
    [Google Scholar]
  73. Borisov VB, Forte E, Siletsky SA, Arese M, Davletshin AI et al. Cytochrome bd protects bacteria against oxidative and nitrosative stress: a potential target for next-generation antimicrobial agents. Biochemistry 2015;80:565–575 [CrossRef]
    [Google Scholar]
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