1887

Abstract

DK1622 possesses two copies of the gene: , which participates in development, and , which is involved in the predatory ability of cells. In this study, we determined that the gene is required for the biosynthesis of the secondary metabolite myxovirescin (TA), which plays essential roles in predation. The -knockout mutant strain was defective in producing a zone of inhibition and displayed decreased killing ability against , while the -knockout mutant strain exhibited little difference from the wild-type strain DK1622. HPLC revealed that deletion of the gene blocked the production of TA, which was present in the -knockout mutant. The addition of exogenous TA rescued the inhibition and killing abilities of the -knockout mutant against . Analysis of GroEL domain-swapping mutants indicated that the C-terminal equatorial domain of GroEL2 was essential for TA production, while the N-terminal equatorial or apical domains of GroEL2 were not sufficient to rescue TA production of the knockout.

Funding
This study was supported by the:
  • National Science Foundation of China (NSFC) Key Program (Award 31130004)
  • NSFC for Distinguished Young Scholars (Award 30825001)
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2014-03-01
2024-12-10
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References

  1. Anscombe F. J., Singh B. N. ( 1948). Limitation of bacteria by micro-predators in soil. Nature 161:140–141 [View Article][PubMed]
    [Google Scholar]
  2. Azem A., Kessel M., Goloubinoff P. ( 1994). Characterization of a functional GroEL14(GroES7)2 chaperonin hetero-oligomer. Science 265:653–656 [View Article][PubMed]
    [Google Scholar]
  3. Berleman J. E., Kirby J. R. ( 2009). Deciphering the hunting strategy of a bacterial wolfpack. FEMS Microbiol Rev 33:942–957 [View Article][PubMed]
    [Google Scholar]
  4. Bittner A. N., Foltz A., Oke V. ( 2007). Only one of five groEL genes is required for viability and successful symbiosis in Sinorhizobium meliloti. J Bacteriol 189:1884–1889 [View Article][PubMed]
    [Google Scholar]
  5. Brocchieri L., Karlin S. ( 2000). Conservation among HSP60 sequences in relation to structure, function, and evolution. Protein Sci 9:476–486 [View Article][PubMed]
    [Google Scholar]
  6. Dworkin M., Kaiser D. ( 1993). Myxobacteria II Washington, DC: American Society for Microbiology;
    [Google Scholar]
  7. Fayet O., Ziegelhoffer T., Georgopoulos C. ( 1989). The groES and groEL heat shock gene products of Escherichia coli are essential for bacterial growth at all temperatures. J Bacteriol 171:1379–1385[PubMed]
    [Google Scholar]
  8. Gaspari F., Paitan Y., Mainini M., Losi D., Ron E. Z., Marinelli F. ( 2005). Myxobacteria isolated in Israel as potential source of new anti-infectives. J Appl Microbiol 98:429–439 [View Article][PubMed]
    [Google Scholar]
  9. George R., Kelly S. M., Price N. C., Erbse A., Fisher M., Lund P. A. ( 2004). Three GroEL homologues from Rhizobium leguminosarum have distinct in vitro properties. Biochem Biophys Res Commun 324:822–828 [View Article][PubMed]
    [Google Scholar]
  10. Gerth K., Irschik H., Reichenbach H., Trowitzsch W. ( 1982). The myxovirescins, a family of antibiotics from Myxococcus virescens (Myxobacterales). J Antibiot (Tokyo) 35:1454–1459 [View Article][PubMed]
    [Google Scholar]
  11. Gerth K., Pradella S., Perlova O., Beyer S., Müller R. ( 2003). Myxobacteria: proficient producers of novel natural products with various biological activities–past and future biotechnological aspects with the focus on the genus Sorangium. J Biotechnol 106:233–253 [View Article][PubMed]
    [Google Scholar]
  12. Goldman B. S., Nierman W. C., Kaiser D., Slater S. C., Durkin A. S., Eisen J. A., Ronning C. M., Barbazuk W. B., Blanchard M. & other authors ( 2006). Evolution of sensory complexity recorded in a myxobacterial genome. Proc Natl Acad Sci U S A 103:15200–15205 [View Article][PubMed]
    [Google Scholar]
  13. Gould P. S., Burgar H. R., Lund P. A. ( 2007). Homologous cpn60 genes in Rhizobium leguminosarum are not functionally equivalent. Cell Stress Chaperones 12:123–131 [View Article][PubMed]
    [Google Scholar]
  14. Goyal K., Qamra R., Mande S. C. ( 2006). Multiple gene duplication and rapid evolution in the groEL gene: functional implications. J Mol Evol 63:781–787 [View Article][PubMed]
    [Google Scholar]
  15. Hodgkin J., Kaiser D. ( 1977). Cell-to-cell stimulation of movement in nonmotile mutants of Myxococcus. Proc Natl Acad Sci U S A 74:2938–2942 [View Article][PubMed]
    [Google Scholar]
  16. Hu Y., Henderson B., Lund P. A., Tormay P., Ahmed M. T., Gurcha S. S., Besra G. S., Coates A. R. ( 2008). A Mycobacterium tuberculosis mutant lacking the groEL homologue cpn60.1 is viable but fails to induce an inflammatory response in animal models of infection. Infect Immun 76:1535–1546 [View Article][PubMed]
    [Google Scholar]
  17. Kashefi K., Hartzell P. L. ( 1995). Genetic suppression and phenotypic masking of a Myxococcus xanthus frzF- defect. Mol Microbiol 15:483–494 [View Article][PubMed]
    [Google Scholar]
  18. Li J., Wang Y., Zhang C. Y., Zhang W. Y., Jiang D. M., Wu Z. H., Liu H., Li Y. Z. ( 2010). Myxococcus xanthus viability depends on groEL supplied by either of two genes, but the paralogs have different functions during heat shock, predation, and development. J Bacteriol 192:1875–1881 [View Article][PubMed]
    [Google Scholar]
  19. Lin Z., Rye H. S. ( 2006). GroEL-mediated protein folding: making the impossible, possible. Crit Rev Biochem Mol Biol 41:211–239 [View Article][PubMed]
    [Google Scholar]
  20. Lund P. A. ( 2001). Microbial molecular chaperones. Adv Microb Physiol 44:93–140 [View Article][PubMed]
    [Google Scholar]
  21. Lund P. A. ( 2009). Multiple chaperonins in bacteria–why so many. FEMS Microbiol Rev 33:785–800 [View Article][PubMed]
    [Google Scholar]
  22. McBride M. J., Zusman D. R. ( 1996). Behavioral analysis of single cells of Myxococcus xanthus in response to prey cells of Escherichia coli. FEMS Microbiol Lett 137:227–231 [View Article][PubMed]
    [Google Scholar]
  23. Ojha A., Anand M., Bhatt A., Kremer L., Jacobs W. R. Jr, Hatfull G. F. ( 2005). GroEL1: a dedicated chaperone involved in mycolic acid biosynthesis during biofilm formation in mycobacteria. Cell 123:861–873 [View Article][PubMed]
    [Google Scholar]
  24. Radford S. E. ( 2006). GroEL: more than just a folding cage. Cell 125:831–833 [View Article][PubMed]
    [Google Scholar]
  25. Ranson N. A., White H. E., Saibil H. R. ( 1998). Chaperonins. Biochem J 333:233–242[PubMed]
    [Google Scholar]
  26. Rodríguez-Quiñones F., Maguire M., Wallington E. J., Gould P. S., Yerko V., Downie J. A., Lund P. A. ( 2005). Two of the three groEL homologues in Rhizobium leguminosarum are dispensable for normal growth. Arch Microbiol 183:253–265 [View Article][PubMed]
    [Google Scholar]
  27. Rosenberg E. (editor) ( 1984). Myxobacteria: Development and Cell Interactions New York: Springer; [View Article]
    [Google Scholar]
  28. Shimkets L. J. ( 1990). Social and developmental biology of the myxobacteria. Microbiol Rev 54:473–501[PubMed]
    [Google Scholar]
  29. Simunovic V., Zapp J., Rachid S., Krug D., Meiser P., Müller R. ( 2006). Myxovirescin A biosynthesis is directed by hybrid polyketide synthases/nonribosomal peptide synthetase, 3-hydroxy-3-methylglutaryl-CoA synthases, and trans-acting acyltransferases. ChemBioChem 7:1206–1220 [View Article][PubMed]
    [Google Scholar]
  30. Ueki T., Inouye S., Inouye M. ( 1996). Positive-negative KG cassettes for construction of multi-gene deletions using a single drug marker. Gene 183:153–157 [View Article][PubMed]
    [Google Scholar]
  31. VanBogelen R. A., Acton M. A., Neidhardt F. C. ( 1987). Induction of the heat shock regulon does not produce thermotolerance in Escherichia coli. Genes Dev 1:525–531 [View Article][PubMed]
    [Google Scholar]
  32. Varon M., Cohen S., Rosenberg E. ( 1984). Autocides produced by Myxococcus xanthus. J Bacteriol 160:1146–1150[PubMed]
    [Google Scholar]
  33. Wang Y., Zhang W. Y., Zhang Z., Li J., Li Z. F., Tan Z. G., Zhang T. T., Wu Z. H., Liu H., Li Y. Z. ( 2013). Mechanisms involved in the functional divergence of duplicated GroEL chaperonins in Myxococcus xanthus DK1622. PLoS Genet 9:e1003306 [View Article][PubMed]
    [Google Scholar]
  34. Weissman K. J., Müller R. ( 2009). A brief tour of myxobacterial secondary metabolism. Bioorg Med Chem 17:2121–2136 [View Article][PubMed]
    [Google Scholar]
  35. Weissman K. J., Müller R. ( 2010). Myxobacterial secondary metabolites: bioactivities and modes-of-action. Nat Prod Rep 27:1276–1295 [View Article][PubMed]
    [Google Scholar]
  36. Wenzel S. C., Müller R. ( 2009). Myxobacteria–‘microbial factories’ for the production of bioactive secondary metabolites. Mol Biosyst 5:567–574 [View Article][PubMed]
    [Google Scholar]
  37. Whitworth D. E. ( 2007). Myxobacteria: Multicellularity and Differentiation Washington, DC: American Society for Microbiology;
    [Google Scholar]
  38. Wu S. S., Kaiser D. ( 1996). Markerless deletions of pil genes in Myxococcus xanthus generated by counterselection with the Bacillus subtilis sacB gene. J Bacteriol 178:5817–5821[PubMed]
    [Google Scholar]
  39. Xiao Y., Wei X., Ebright R., Wall D. ( 2011). Antibiotic production by myxobacteria plays a role in predation. J Bacteriol 193:4626–4633 [View Article][PubMed]
    [Google Scholar]
  40. Zhang W., Li Y., Qian G., Wang Y., Chen H., Li Y. Z., Liu F., Shen Y., Du L. ( 2011). Identification and characterization of the anti-methicillin-resistant Staphylococcus aureus WAP-8294A2 biosynthetic gene cluster from Lysobacter enzymogenes OH11. Antimicrob Agents Chemother 55:5581–5589 [View Article][PubMed]
    [Google Scholar]
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