1887

Abstract

The synthesis of methionine is critical for most bacteria. It is known that cellular methionine has a feedback effect on the expression of met genes involved in de novo methionine biosynthesis. Previous studies revealed that Gram-negative bacteria control met gene expression at the transcriptional level by regulator proteins, while most Gram-positive bacteria regulate met genes at post-transcriptional level by RNA regulators (riboregulators) located in the 5′UTR of met genes. However, despite its importance, the methionine biosynthesis pathway in the Gram-negative Xanthomonas genus that includes many important plant pathogens is completely uncharacterized. Here, we address this issue using the crucifer black rot pathogen Xanthomonas campestris pv. campestris (Xcc), a model bacterium in microbe–plant interaction studies. The work identified an operon (met) involved in de novo methionine biosynthesis in Xcc. Disruption of the operon resulted in defective growth in methionine-limited media and in planta. Western blot analysis revealed that the expression of the operon is dependent on methionine levels. Further molecular analyses demonstrated that the 5′UTR, but not the promoter of the operon, is involved in feedback regulation on operon expression in response to methionine availability, providing an example of a Gram-negative bacterium utilizing a 5′UTR region to control the expression of the genes involved in methionine biosynthesis.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000690
2018-07-19
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/164/9/1146.html?itemId=/content/journal/micro/10.1099/mic.0.000690&mimeType=html&fmt=ahah

References

  1. Serganov A, Patel DJ. Amino acid recognition and gene regulation by riboswitches. Biochim Biophys Acta 2009; 1789:592–611 [View Article][PubMed]
    [Google Scholar]
  2. Fontecave M, Atta M, Mulliez E. S-adenosylmethionine: nothing goes to waste. Trends Biochem Sci 2004; 29:243–249 [View Article][PubMed]
    [Google Scholar]
  3. McCutcheon JP, Moran NA. Extreme genome reduction in symbiotic bacteria. Nat Rev Microbiol 2012; 10:13–26 [View Article]
    [Google Scholar]
  4. Ferla MP, Patrick WM. Bacterial methionine biosynthesis. Microbiology 2014; 160:1571–1584 [View Article][PubMed]
    [Google Scholar]
  5. Viola RE. The central enzymes of the aspartate family of amino acid biosynthesis. Acc Chem Res 2001; 34:339–349 [View Article][PubMed]
    [Google Scholar]
  6. Sherwood AV, Henkin TM. Riboswitch-mediated gene regulation: novel RNA architectures dictate gene expression responses. Annu Rev Microbiol 2016; 70:361–374 [View Article][PubMed]
    [Google Scholar]
  7. Breaker RR. Prospects for riboswitch discovery and analysis. Mol Cell 2011; 43:867–879 [View Article][PubMed]
    [Google Scholar]
  8. Rodionov DA, Vitreschak AG, Mironov AA, Gelfand MS. Comparative genomics of the methionine metabolism in Gram-positive bacteria: a variety of regulatory systems. Nucleic Acids Res 2004; 32:3340–3353 [View Article][PubMed]
    [Google Scholar]
  9. Gutiérrez-Preciado A, Henkin TM, Grundy FJ, Yanofsky C, Merino E. Biochemical features and functional implications of the RNA-based T-box regulatory mechanism. Microbiol Mol Biol Rev 2009; 73:36–61 [View Article][PubMed]
    [Google Scholar]
  10. Leyn SA, Suvorova IA, Kholina TD, Sherstneva SS, Novichkov PS et al. Comparative genomics of transcriptional regulation of methionine metabolism in Proteobacteria. PLoS One 2014; 9:e113714 [View Article][PubMed]
    [Google Scholar]
  11. Cubitt MF, Hedley PE, Williamson NR, Morris JA, Campbell E et al. A metabolic regulator modulates virulence and quorum sensing signal production in Pectobacterium atrosepticum. Mol Plant Microbe Interact 2013; 26:356–366 [View Article][PubMed]
    [Google Scholar]
  12. Andersen GL, Beattie GA, Lindow SE. Molecular characterization and sequence of a methionine biosynthetic locus from Pseudomonas syringae. J Bacteriol 1998; 180:4497–4507[PubMed]
    [Google Scholar]
  13. Plener L, Boistard P, González A, Boucher C, Genin S. Metabolic adaptation of Ralstonia solanacearum during plant infection: a methionine biosynthesis case study. PLoS One 2012; 7:e36877 [View Article][PubMed]
    [Google Scholar]
  14. Mansfield J, Genin S, Magori S, Citovsky V, Sriariyanum M et al. Top 10 plant pathogenic bacteria in molecular plant pathology. Mol Plant Pathol 2012; 13:614–629 [View Article][PubMed]
    [Google Scholar]
  15. Li L, Li RF, Ming ZH, Lu GT, Tang JL. Identification of a novel type III secretion-associated outer membrane-bound protein from Xanthomonas campestris pv. campestris. Sci Rep 2017; 7:42724 [View Article][PubMed]
    [Google Scholar]
  16. Windgassen M, Urban A, Jaeger KE. Rapid gene inactivation in Pseudomonas aeruginosa. FEMS Microbiol Lett 2000; 193:201–205 [View Article][PubMed]
    [Google Scholar]
  17. Patey G, Qi Z, Bourg G, Baron C, O'Callaghan D. Swapping of periplasmic domains between Brucella suis VirB8 and a pSB102 VirB8 homologue allows heterologous complementation. Infect Immun 2006; 74:4945–4949 [View Article][PubMed]
    [Google Scholar]
  18. Jefferson RA, Burgess SM, Hirsh D. beta-glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci USA 1986; 83:8447–8451 [View Article][PubMed]
    [Google Scholar]
  19. Qian W, Jia Y, Ren SX, He YQ, Feng JX et al. Comparative and functional genomic analyses of the pathogenicity of phytopathogen Xanthomonas campestris pv. campestris. Genome Res 2005; 15:757–767 [View Article][PubMed]
    [Google Scholar]
  20. Weissbach H, Brot N. Regulation of methionine synthesis in Escherichia coli. Mol Microbiol 1991; 5:1593–1597 [View Article][PubMed]
    [Google Scholar]
  21. Urbanowski ML, Stauffer LT, Plamann LS, Stauffer GV. A new methionine locus, metR, that encodes a trans-acting protein required for activation of metE and metH in Escherichia coli and Salmonella typhimurium. J Bacteriol 1987; 169:1391–1397 [View Article][PubMed]
    [Google Scholar]
  22. Wang JX, Lee ER, Morales DR, Lim J, Breaker RR. Riboswitches that sense S-adenosylhomocysteine and activate genes involved in coenzyme recycling. Mol Cell 2008; 29:691–702 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000690
Loading
/content/journal/micro/10.1099/mic.0.000690
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