1887

Abstract

Polyamines (PAs) are ubiquitous polycations derived from basic -amino acids whose physiological roles are still being defined. Their biosynthesis and functions in nitrogen-fixing rhizobia such as have not been extensively investigated. Thin layer chromatographic and mass spectrometric analyses showed that Rm8530 produces the PAs, putrescine (Put), spermidine (Spd) and homospermidine (HSpd), in their free forms and norspermidine (NSpd) in a form bound to macromolecules. The genome encodes two putative ornithine decarboxylases (ODC) for Put synthesis. Activity assays with the purified enzymes showed that ODC2 (SMc02983) decarboxylates both ornithine and lysine. ODC1 (SMa0680) decarboxylates only ornithine. An mutant was similar to the wild-type in ODC activity, PA production and growth. In comparison to the wild-type, an mutant had 45 % as much ODC activity and its growth rates were reduced by 42, 14 and 44 % under non-stress, salt stress or acid stress conditions, respectively. The mutant produced only trace levels of Put, Spd and HSpd. Wild-type phenotypes were restored when the mutant was grown in cultures supplemented with 1 mM Put or Spd or when the gene was introduced gene expression was increased under acid stress and reduced under salt stress and with exogenous Put or Spd. An double mutant had phenotypes similar to the mutant. These results indicate that ODC2 is the major enzyme for Put synthesis in and that PAs are required for normal growth .

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000615
2018-04-01
2020-11-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/164/4/600.html?itemId=/content/journal/micro/10.1099/mic.0.000615&mimeType=html&fmt=ahah

References

  1. Tabor CW, Tabor H. Polyamines. Annu Rev Biochem 1984;53:749–790 [CrossRef][PubMed]
    [Google Scholar]
  2. Hamana K, Matsuzaki S. Polyamines as a chemotaxonomic marker in bacterial systematics. Crit Rev Microbiol 1992;18:261–283 [CrossRef][PubMed]
    [Google Scholar]
  3. Algranati ID. Polyamine metabolism in Trypanosoma cruzi: studies on the expression and regulation of heterologous genes involved in polyamine biosynthesis. Amino Acids 2010;38:645–651 [CrossRef][PubMed]
    [Google Scholar]
  4. Miller-Fleming L, Olin-Sandoval V, Campbell K, Ralser M. Remaining mysteries of molecular biology: the role of polyamines in the cell. J Mol Biol 2015;427:3389–3406 [CrossRef][PubMed]
    [Google Scholar]
  5. Michael AJ. Polyamines in eukaryotes, bacteria, and archaea. J Biol Chem 2016;291:14896–14903 [CrossRef][PubMed]
    [Google Scholar]
  6. Soksawatmaekhin W, Kuraishi A, Sakata K, Kashiwagi K, Igarashi K. Excretion and uptake of cadaverine by CadB and its physiological functions in Escherichia coli. Mol Microbiol 2004;51:1401–1412 [CrossRef][PubMed]
    [Google Scholar]
  7. Sturgill G, Rather PN. Evidence that putrescine acts as an extracellular signal required for swarming in Proteus mirabilis. Mol Microbiol 2004;51:437–446 [CrossRef][PubMed]
    [Google Scholar]
  8. Shah P, Swiatlo E. A multifaceted role for polyamines in bacterial pathogens. Mol Microbiol 2008;68:4–16 [CrossRef][PubMed]
    [Google Scholar]
  9. Lee J, Sperandio V, Frantz DE, Longgood J, Camilli A et al. An alternative polyamine biosynthetic pathway is widespread in bacteria and essential for biofilm formation in Vibrio cholerae. J Biol Chem 2009;284:9899–9907 [CrossRef][PubMed]
    [Google Scholar]
  10. Cockerell SR, Rutkovsky AC, Zayner JP, Cooper RE, Porter LR et al. Vibrio cholerae NspS, a homologue of ABC-type periplasmic solute binding proteins, facilitates transduction of polyamine signals independent of their transport. Microbiology 2014;160:832–843 [CrossRef][PubMed]
    [Google Scholar]
  11. Kim SH, Wang Y, Khomutov M, Khomutov A, Fuqua C et al. The essential role of spermidine in growth of Agrobacterium tumefaciens Is determined by the 1,3-diaminopropane moiety. ACS Chem Biol 2016;11:491–499 [CrossRef][PubMed]
    [Google Scholar]
  12. López-Gómez M, Hidalgo-Castellanos J, Lluch C, Herrera-Cervera JA. 24-Epibrassinolide ameliorates salt stress effects in the symbiosis Medicago truncatula-Sinorhizobium meliloti and regulates the nodulation in cross-talk with polyamines. Plant Physiol Biochem 2016;108:212–221 [CrossRef][PubMed]
    [Google Scholar]
  13. Lucas PM. Ornithine and lysine decarboxylation in bacteria. In D'Mello FJ. (editor) Handbook of Microbial Metabolism of Amino Acids Wallingford, Oxfordshire: CAB International; 2017; pp.116–127[Crossref]
    [Google Scholar]
  14. Dunn MF. Rhizobial amino acid metabolism: polyamine biosynthesis and functions. In D'Mello FJ. (editor) Handbook of Microbial Metabolism of Amino Acids Wallingford, Oxfordshire: CAB International; 2017; pp.352–370[Crossref]
    [Google Scholar]
  15. Hamana K, Minamisawa K, Matsuzaki S. Polyamines in Rhizobium, Bradyrhizobium, Azorhizobium and Agrobacterium. FEMS Microbiol Lett 1990;71:71–76 [CrossRef]
    [Google Scholar]
  16. Hamana K, Sakamoto A, Tachiyanagi S, Terauchi E, Takeuchi M. Polyamine profiles of some members of the alpha subclass of the class Proteobacteria: Polyamine analysis of twenty recently described genera. Microbiol Cult Coll 2003;19:13–21
    [Google Scholar]
  17. Vassileva V, Ignatov G. Polyamine-induced changes in symbiotic parameters of the Galega orientalis-Rhizobium galegae nitrogen-fixing system. Plant Soil 1999;210:83–91 [CrossRef]
    [Google Scholar]
  18. Fujihara S, Yoneyama T. Effects of pH and osmotic stress on cellular polyamine contents in the soybean rhizobia Rhizobium fredii P220 and Bradyrhizobium japonicum A1017. Appl Environ Microbiol 1993;59:1104–1109[PubMed]
    [Google Scholar]
  19. López-Gómez M, Hidalgo-Castellanos J, Iribarne C, Lluch C. Proline accumulation has prevalence over polyamines in nodules of Medicago sativa in symbiosis with Sinorhizobium meliloti during the initial response to salinity. Plant Soil 2014;374:149–159 [CrossRef]
    [Google Scholar]
  20. Palma F, López-Gómez M, Tejera NA, Lluch C. Involvement of abscisic acid in the response of Medicago sativa plants in symbiosis with Sinorhizobium meliloti to salinity. Plant Sci 2014;223:16–24 [CrossRef][PubMed]
    [Google Scholar]
  21. López-Gómez M, Hidalgo-Castellanos J, Muñoz-Sánchez JR, Marín-Peña AJ, Lluch C et al. Polyamines contribute to salinity tolerance in the symbiosis Medicago truncatula-Sinorhizobium meliloti by preventing oxidative damage. Plant Physiol Biochem 2017;116:9–17 [CrossRef][PubMed]
    [Google Scholar]
  22. Braeken K, Daniels R, Vos K, Fauvart M, Bachaspatimayum D et al. Genetic determinants of swarming in Rhizobium etli. Microb Ecol 2008;55:54–64 [CrossRef][PubMed]
    [Google Scholar]
  23. Shaw FL. From prediction to function: Polyamine biosynthesisand formate metabolism in the α- and ε-Proteobacteria. Ph.D Thesis England: Institute of Food Research, Norwich Research Park; 2011
    [Google Scholar]
  24. López-Gómez M, Cobos-Porras L, Prell J, Lluch C. Homospermidine synthase contributes to salt tolerance in free-living Rhizobium tropici and in symbiosis with Phaseolus vulgaris. Plant Soil 2016;404:413–425 [CrossRef]
    [Google Scholar]
  25. Wang Y, Kim SH, Natarajan R, Heindl JE, Bruger EL et al. Spermidine inversely influences surface interactions and planktonic growth in Agrobacterium tumefaciens. J Bacteriol 2016;198:2682–2691 [CrossRef][PubMed]
    [Google Scholar]
  26. Dunn MF. Key roles of microsymbiont amino acid metabolism in rhizobia-legume interactions. Crit Rev Microbiol 2015;41:411–451 [CrossRef][PubMed]
    [Google Scholar]
  27. Hernández VM, Girard L, Hernández-Lucas I, Vázquez A, Ortíz-Ortíz C et al. Genetic and biochemical characterization of arginine biosynthesis in Sinorhizobium meliloti 1021. Microbiology 2015;161:1671–1682 [CrossRef][PubMed]
    [Google Scholar]
  28. Dunn MF, Araíza G, Cevallos MA, Mora J. Regulation of pyruvate carboxylase in Rhizobium etli. FEMS Microbiol Lett 1997;157:301–306 [CrossRef][PubMed]
    [Google Scholar]
  29. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory Press; 1989
    [Google Scholar]
  30. Landeta C, Dávalos A, Cevallos , Geiger O, Brom S et al. Plasmids with a chromosome-like role in Rhizobia. J Bacteriol 2011;193:1317–1326 [CrossRef][PubMed]
    [Google Scholar]
  31. Romano A, Trip H, Lolkema JS, Lucas PM. Three-component lysine/ornithine decarboxylation system in Lactobacillus saerimneri 30a. J Bacteriol 2013;195:1249–1254 [CrossRef][PubMed]
    [Google Scholar]
  32. Girard L, Brom S, Dávalos A, López O, Soberón M et al. Differential regulation of fixN-reiterated genes in Rhizobium etli by a novel fixL-fixK cascade. Mol Plant Microbe Interact 2000;13:1283–1292 [CrossRef][PubMed]
    [Google Scholar]
  33. Goldschmidt MC, Lockhart BM. Rapid methods for determining decarboxylase activity: arginine decarboxylase. Appl Microbiol 1971;22:350–357[PubMed]
    [Google Scholar]
  34. Ngo TT, Brillhart KL, Davis RH, Wong RC, Bovaird JH et al. Spectrophotometric assay for ornithine decarboxylase. Anal Biochem 1987;160:290–293 [CrossRef][PubMed]
    [Google Scholar]
  35. Phan AP, Ngo TT, Lenhoff HM. Spectrophotometric assay for lysine decarboxylase. Anal Biochem 1982;120:193–197 [CrossRef][PubMed]
    [Google Scholar]
  36. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–254 [CrossRef][PubMed]
    [Google Scholar]
  37. Schlüter JP, Reinkensmeier J, Barnett MJ, Lang C, Krol E et al. Global mapping of transcription start sites and promoter motifs in the symbiotic α-proteobacterium Sinorhizobium meliloti 1021. BMC Genomics 2013;14:156 [CrossRef][PubMed]
    [Google Scholar]
  38. Slocum RD, Flores HE, Galston AW, Weinstein LH. Improved method for HPLC analysis of polyamines, agmatine and aromatic monoamines in plant tissue. Plant Physiol 1989;89:512–517 [CrossRef][PubMed]
    [Google Scholar]
  39. Pedrol N, Tiburcio AF. Polyamines determination by TLC and HPLC. In Roger MJR. (editor) Handbook of Plant Ecophysiology Techniques The Netherlands: Kluwer Academic Publishers; 2001; pp.335–363
    [Google Scholar]
  40. Pellock BJ, Teplitski M, Boinay RP, Bauer WD, Walker GC. A LuxR homolog controls production of symbiotically active extracellular polysaccharide II by Sinorhizobium meliloti. J Bacteriol 2002;184:5067–5076 [CrossRef][PubMed]
    [Google Scholar]
  41. Paulino EM. Búsqueda del gen que codifica para la enzima N-acetilglutamato sintasa por mutagénesis. Bachelors Thesis Mexico: Universidad Autónoma del Estado de Morelos; 2013
    [Google Scholar]
  42. Domínguez-Ferreras A, Pérez-Arnedo R, Becker A, Olivares J, Soto MJ et al. Transcriptome profiling reveals the importance of plasmid pSymB for osmoadaptation of Sinorhizobium meliloti. J Bacteriol 2006;188:7617–7625 [CrossRef][PubMed]
    [Google Scholar]
  43. Kovach ME, Elzer PH, Hill DS, 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][PubMed]
    [Google Scholar]
  44. Schäfer A, Tauch A, Jäger W, Kalinowski J, Thierbach G et al. Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 1994;145:69–73 [CrossRef][PubMed]
    [Google Scholar]
  45. Martinez-Salazar JM, Romero D. Role of the ruvB gene in homologous and homeologous recombination in Rhizobium etli. Gene 2000;243:125–131 [CrossRef][PubMed]
    [Google Scholar]
  46. Figurski DH, Helinski DR. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 1979;76:1648–1652 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000615
Loading
/content/journal/micro/10.1099/mic.0.000615
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF

Most cited this month 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