1887

Abstract

The soil bacterium , a nitrogen-fixing symbiont of legume plants, is exposed to numerous stress conditions in nature, some of which cause the formation of harmful DNA double-strand breaks (DSBs). In particular, the reactive oxygen species (ROS) and the reactive nitrogen species (RNS) produced during symbiosis, and the desiccation occurring in dry soils, are conditions which induce DSBs. Two major systems of DSB repair are known in : homologous recombination (HR) and non-homologous end-joining (NHEJ). However, their role in the resistance to ROS, RNS and desiccation has never been examined in this bacterial species, and the importance of DSB repair in the symbiotic interaction has not been properly evaluated. Here, we constructed strains deficient in HR (by deleting the gene) or in NHEJ (by deleting the four genes) or both. Interestingly, we observed that and/or genes are involved in resistance to ROS and RNS. Nevertheless, an strain deficient in both HR and NHEJ was not altered in its ability to establish and maintain an efficient nitrogen-fixing symbiosis with , showing that rhizobial DSB repair is not essential for this process. This result suggests either that DSB formation in is efficiently prevented during symbiosis or that DSBs are not detrimental for symbiosis efficiency. In contrast, we found for the first time that both and genes are involved in resistance to desiccation, suggesting that DSB repair could be important for rhizobium persistence in the soil.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000400
2017-03-01
2024-03-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/163/3/333.html?itemId=/content/journal/micro/10.1099/mic.0.000400&mimeType=html&fmt=ahah

References

  1. Bennett CB, Lewis AL, Baldwin KK, Resnick MA. Lethality induced by a single site-specific double-strand break in a dispensable yeast plasmid. Proc Natl Acad Sci USA 1993; 90:5613–5617 [View Article][PubMed]
    [Google Scholar]
  2. Michel B, Leach D. Homologous recombination – enzymes and pathways. EcoSal Plus 2012 doi:10.1128/ecosalplus.7.2.7 [View Article][PubMed]
    [Google Scholar]
  3. Janion C. Inducible SOS response system of DNA repair and mutagenesis in Escherichia coli. Int J Biol Sci 2008; 4:338–344 [View Article][PubMed]
    [Google Scholar]
  4. Persky NS, Lovett ST. Mechanisms of recombination: lessons from E. coli. Crit Rev Biochem Mol Biol 2008; 43:347–370 [View Article][PubMed]
    [Google Scholar]
  5. Aravind L, Koonin EV. Prokaryotic homologs of the eukaryotic DNA-end-binding protein Ku, novel domains in the Ku protein and prediction of a prokaryotic double-strand break repair system. Genome Res 2001; 11:1365–1374 [View Article][PubMed]
    [Google Scholar]
  6. Mcgovern S, Baconnais S, Roblin P, Nicolas P, Drevet P et al. C-terminal region of bacterial Ku controls DNA bridging, DNA threading and recruitment of DNA ligase D for double strand breaks repair. Nucleic Acids Res 20164785–4806 [View Article][PubMed]
    [Google Scholar]
  7. Weller GR, Kysela B, Roy R, Tonkin LM, Scanlan E et al. Identification of a DNA nonhomologous end-joining complex in bacteria. Science 2002; 297:1686–1689 [View Article][PubMed]
    [Google Scholar]
  8. Pitcher RS, Brissett NC, Doherty AJ. Nonhomologous end-joining in bacteria: a microbial perspective. Annu Rev Microbiol 2007; 61:259–282 [View Article][PubMed]
    [Google Scholar]
  9. Shuman S, Glickman MS. Bacterial DNA repair by non-homologous end joining. Nat Rev Microbiol 2007; 5:852–861 [View Article][PubMed]
    [Google Scholar]
  10. Pitcher RS, Green AJ, Brzostek A, Korycka-Machala M, Dziadek J et al. NHEJ protects mycobacteria in stationary phase against the harmful effects of desiccation. DNA Repair 2007; 6:1271–1276 [View Article][PubMed]
    [Google Scholar]
  11. Stephanou NC, Gao F, Bongiorno P, Ehrt S, Schnappinger D et al. Mycobacterial nonhomologous end joining mediates mutagenic repair of chromosomal double-strand DNA breaks. J Bacteriol 2007; 189:5237–5246 [View Article][PubMed]
    [Google Scholar]
  12. Brzostek A, Szulc I, Klink M, Brzezinska M, Sulowska Z et al. Either non-homologous ends joining or homologous recombination is required to repair double-strand breaks in the genome of macrophage-internalized Mycobacterium tuberculosis. PLoS One 2014; 9:e92799 [View Article][PubMed]
    [Google Scholar]
  13. Wang ST, Setlow B, Conlon EM, Lyon JL, Imamura D et al. The forespore line of gene expression in Bacillus subtilis. J Mol Biol 2006; 358:16–37 [View Article][PubMed]
    [Google Scholar]
  14. Better M, Helinski DR. Isolation and characterization of the recA gene of Rhizobium meliloti. J Bacteriol 1983; 155:311–316[PubMed]
    [Google Scholar]
  15. Kobayashi H, Simmons LA, Yuan DS, Broughton WJ, Walker GC. Multiple ku orthologues mediate DNA non-homologous end-joining in the free-living form and during chronic infection of Sinorhizobium meliloti. Mol Microbiol 2008; 67:350–363 [View Article][PubMed]
    [Google Scholar]
  16. Selbitschka W, Arnold W, Priefer UB, Rottschäfer T, Schmidt M et al. Characterization of recA genes and recA mutants of Rhizobium meliloti and Rhizobium leguminosarum biovar viciae. Mol Gen Genet 1991; 229:86–95 [View Article][PubMed]
    [Google Scholar]
  17. Udvardi M, Poole PS. Transport and metabolism in legume-rhizobia symbioses. Annu Rev Plant Biol 2013; 64:781–805 [View Article][PubMed]
    [Google Scholar]
  18. Zahran HH. Rhizobium-legume symbiosis and nitrogen fixation under severe conditions and in an arid climate. Microbiol Mol Biol Rev 1999; 63:968–989[PubMed]
    [Google Scholar]
  19. Vriezen JA, de Bruijn FJ, Nüsslein K. Responses of rhizobia to desiccation in relation to osmotic stress, oxygen, and temperature. Appl Environ Microbiol 2007; 73:3451–3459 [View Article][PubMed]
    [Google Scholar]
  20. Baudouin E, Pieuchot L, Engler G, Pauly N, Puppo A. Nitric oxide is formed in Medicago truncatulaSinorhizobium meliloti functional nodules. Mol Plant Microbe Interact 2006; 19:970–975 [View Article][PubMed]
    [Google Scholar]
  21. del Giudice J, Cam Y, Damiani I, Fung-Chat F, Meilhoc E et al. Nitric oxide is required for an optimal establishment of the Medicago truncatulaSinorhizobium meliloti symbiosis. New Phytol 2011; 191:405–417 [View Article][PubMed]
    [Google Scholar]
  22. Rubio MC, James EK, Clemente MR, Bucciarelli B, Fedorova M et al. Localization of superoxide dismutases and hydrogen peroxide in legume root nodules. Mol Plant Microbe Interact 2004; 17:1294–1305 [View Article][PubMed]
    [Google Scholar]
  23. Santos R, Hérouart D, Puppo A, Touati D. Critical protective role of bacterial superoxide dismutase in rhizobium-legume symbiosis. Mol Microbiol 2000; 38:750–759 [View Article][PubMed]
    [Google Scholar]
  24. Driessens N, Versteyhe S, Ghaddhab C, Burniat A, de Deken X et al. Hydrogen peroxide induces DNA single- and double-strand breaks in thyroid cells and is therefore a potential mutagen for this organ. Endocr Relat Cancer 2009; 16:845–856 [View Article][PubMed]
    [Google Scholar]
  25. Tamir S, Burney S, Tannenbaum SR. DNA damage by nitric oxide. Chem Res Toxicol 1996; 9:821–827 [View Article][PubMed]
    [Google Scholar]
  26. Ferri L, Gori A, Biondi EG, Mengoni A, Bazzicalupo M. Plasmid electroporation of Sinorhizobium strains: the role of the restriction gene hsdR in type strain Rm1021. Plasmid 2010; 63:128–135 [View Article][PubMed]
    [Google Scholar]
  27. Humann JL, Ziemkiewicz HT, Yurgel SN, Kahn ML. Regulatory and DNA repair genes contribute to the desiccation resistance of Sinorhizobium meliloti Rm1021. Appl Environ Microbiol 2009; 75:446–453 [View Article][PubMed]
    [Google Scholar]
  28. Berrabah F, Bourcy M, Cayrel A, Eschstruth A, Mondy S et al. Growth conditions determine the DNF2 requirement for symbiosis. PLoS One 2014; 9:e91866 [View Article][PubMed]
    [Google Scholar]
  29. Fahraeus G. The infection of clover root hairs by nodule bacteria studied by a simple glass slide technique. J Gen Microbiol 1957; 16:374–381 [View Article][PubMed]
    [Google Scholar]
  30. Meilhoc E, Cam Y, Skapski A, Bruand C. The response to nitric oxide of the nitrogen-fixing symbiont Sinorhizobium meliloti. Mol Plant Microbe Interact 2010; 23:748–759 [View Article][PubMed]
    [Google Scholar]
  31. Graham PM, Li JZ, Dou X, Zhu H, Misra HP et al. Protection against peroxynitrite-induced DNA damage by mesalamine: implications for anti-inflammation and anti-cancer activity. Mol Cell Biochem 2013; 378:291–298 [View Article][PubMed]
    [Google Scholar]
  32. Mattimore V, Battista JR. Radioresistance of Deinococcus radiodurans: functions necessary to survive ionizing radiation are also necessary to survive prolonged desiccation. J Bacteriol 1996; 178:633–637 [View Article][PubMed]
    [Google Scholar]
  33. Potts M. Desiccation tolerance of prokaryotes. Microbiol Rev 1994; 58:755–805[PubMed]
    [Google Scholar]
  34. Jones KM, Kobayashi H, Davies BW, Taga ME, Walker GC. How rhizobial symbionts invade plants: the Sinorhizobium-Medicago model. Nat Rev Microbiol 2007; 5:619–633 [View Article][PubMed]
    [Google Scholar]
  35. Boscari A, Meilhoc E, Castella C, Bruand C, Puppo A et al. Which role for nitric oxide in symbiotic N2-fixing nodules: toxic by-product or useful signaling/metabolic intermediate?. Front Plant Sci 2013; 4: [View Article][PubMed]
    [Google Scholar]
  36. Puppo A, Pauly N, Boscari A, Mandon K, Brouquisse R. Hydrogen peroxide and nitric oxide: key regulators of the legume-rhizobium and mycorrhizal symbioses. Antioxid Redox Signal 2013; 18:2202–2219 [View Article][PubMed]
    [Google Scholar]
  37. Sallet E, Roux B, Sauviac L, Jardinaud M-F, Carrère S et al. Next-generation annotation of prokaryotic genomes with EuGene-P: application to Sinorhizobium meliloti 2011. DNA Res 2013; 20:339–354 [View Article][PubMed]
    [Google Scholar]
  38. Cam Y, Pierre O, Boncompagni E, Hérouart D, Meilhoc E et al. Nitric oxide (NO): a key player in the senescence of Medicago truncatula root nodules. New Phytol 2012; 196:548–560 [View Article][PubMed]
    [Google Scholar]
  39. Jamet A, Sigaud S, van de Sype G, Puppo A, Hérouart D. Expression of the bacterial catalase genes during Sinorhizobium meliloti-Medicago sativa symbiosis and their crucial role during the infection process. Mol Plant Microbe Interact 2003; 16:217–225 [View Article][PubMed]
    [Google Scholar]
  40. Jamet A, Kiss E, Batut J, Puppo A, Hérouart D. The katA catalase gene is regulated by OxyR in both free-living and symbiotic Sinorhizobium meliloti. J Bacteriol 2005; 187:376–381 [View Article][PubMed]
    [Google Scholar]
  41. Meilhoc E, Blanquet P, Cam Y, Bruand C. Control of NO level in rhizobium-legume root nodules. Plant Signal Behav 2013; 8:e25923 [View Article]
    [Google Scholar]
  42. Sigaud S, Becquet V, Frendo P, Puppo A, Hérouart D. Differential regulation of two divergent Sinorhizobium meliloti genes for HPII-like catalases during free-living growth and protective role of both catalases during symbiosis. J Bacteriol 1999; 181:2634–2639[PubMed]
    [Google Scholar]
  43. Alunni B, Gourion B. Terminal bacteroid differentiation in the legume-rhizobium symbiosis: nodule-specific cysteine-rich peptides and beyond. New Phytol 2016; 211:411–417 [View Article][PubMed]
    [Google Scholar]
  44. Aranda J, Bardina C, Beceiro A, Rumbo S, Cabral MP et al. Acinetobacter baumannii RecA protein in repair of DNA damage, antimicrobial resistance, general stress response, and virulence. J Bacteriol 2011; 193:3740–3747 [View Article][PubMed]
    [Google Scholar]
  45. Vlašić I, Mertens R, Seco EM, Carrasco B, Ayora S et al. Bacillus subtilis RecA and its accessory factors, RecF, RecO, RecR and RecX, are required for spore resistance to DNA double-strand break. Nucleic Acids Res 2014; 42:2295–2307 [View Article][PubMed]
    [Google Scholar]
  46. Moeller R, Stackebrandt E, Reitz G, Berger T, Rettberg P et al. Role of DNA repair by nonhomologous-end joining in Bacillus subtilis spore resistance to extreme dryness, mono- and polychromatic UV, and ionizing radiation. J Bacteriol 2007; 189:3306–3311 [View Article][PubMed]
    [Google Scholar]
  47. Cytryn EJ, Sangurdekar DP, Streeter JG, Franck WL, Chang WS et al. Transcriptional and physiological responses of Bradyrhizobium japonicum to desiccation-induced stress. J Bacteriol 2007; 189:6751–6762 [View Article][PubMed]
    [Google Scholar]
  48. Pobigaylo N, Wetter D, Szymczak S, Schiller U, Kurtz S et al. Construction of a large signature-tagged mini-Tn5 transposon library and its application to mutagenesis of Sinorhizobium meliloti. Appl Environ Microbiol 2006; 72:4329–4337 [View Article][PubMed]
    [Google Scholar]
  49. Quandt J, Hynes MF. Versatile suicide vectors which allow direct selection for gene replacement in Gram-negative bacteria. Gene 1993; 127:15–21 [View Article][PubMed]
    [Google Scholar]
  50. Prentki P, Krisch HM. In vitro insertional mutagenesis with a selectable DNA fragment. Gene 1984; 29:303–313 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000400
Loading
/content/journal/micro/10.1099/mic.0.000400
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
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