1887

Abstract

Laboratory strains and natural isolates of differ in their level of stress resistance due to strain variation in the level of the sigma factor (or RpoS), the transcriptional master controller of the general stress response. We found that the high level of RpoS in one laboratory strain (MC4100) was partially dependent on an elevated basal level of ppGpp, an alarmone responding to stress and starvation. The elevated ppGpp was caused by two mutations in , a gene associated with ppGpp synthesis and degradation. The nature of the allele influenced the level of ppGpp in both MC4100 and another commonly used K-12 strain, MG1655. Introduction of the mutation into MG1655 also resulted in an increased level of RpoS, but the amount of RpoS was lower in MG1655 than in MC4100 with either the wild-type or mutant allele. In both MC4100 and MG1655, high ppGpp concentration increased RpoS levels, which in turn reduced growth with poor carbon sources like acetate. The growth inhibition resulting from elevated ppGpp was relieved by mutations. The extent of the growth inhibition by ppGpp, as well as the magnitude of the relief by mutations, differed between MG1655 and MC4100. These results together suggest that mutations represent one of several polymorphisms influencing the strain variation of RpoS levels. Stress resistance was higher in strains with the mutation, which is consistent with the conclusion that microevolution affecting either or both ppGpp and RpoS can reset the balance between self-protection and nutritional capability, the SPANC balance, in individual strains of

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2008/018457-0
2008-09-01
2019-10-23
Loading full text...

Full text loading...

/deliver/fulltext/micro/154/9/2887.html?itemId=/content/journal/micro/10.1099/mic.0.2008/018457-0&mimeType=html&fmt=ahah

References

  1. Atlung, T., Nielsen, H. V. & Hansen, F. G. ( 2002; ). Characterisation of the allelic variation in the rpoS gene in thirteen K-12 and six other non-pathogenic Escherichia coli strains. Mol Genet Genomics 266, 873–881.[CrossRef]
    [Google Scholar]
  2. Bohannon, D. E., Connell, N., Keener, J., Tormo, A., Espinosa-Urgel, M., Zambrano, M. M. & Kolter, R. ( 1991; ). Stationary-phase-inducible “gearbox” promoters: differential effects of katF mutations and role of sigma 70. J Bacteriol 173, 4482–4492.
    [Google Scholar]
  3. Bougdour, A. & Gottesman, S. ( 2007; ). ppGpp regulation of RpoS degradation via anti-adaptor protein IraP. Proc Natl Acad Sci U S A 104, 12896–12901.[CrossRef]
    [Google Scholar]
  4. Braeken, K., Moris, M., Daniels, R., Vanderleyden, J. & Michiels, J. ( 2006; ). New horizons for (p)ppGpp in bacterial and plant physiology. Trends Microbiol 14, 45–54.[CrossRef]
    [Google Scholar]
  5. Brown, L., Gentry, D., Elliott, T. & Cashel, M. ( 2002; ). DksA affects ppGpp induction of RpoS at a translational level. J Bacteriol 184, 4455–4465.[CrossRef]
    [Google Scholar]
  6. Casadaban, M. J. ( 1976; ). Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage Lambda and Mu. J Mol Biol 104, 541–555.[CrossRef]
    [Google Scholar]
  7. Cashel, M. & Gallant, J. ( 1969; ). Two compounds implicated in the function of the RC gene of Escherichia coli. Nature 221, 838–841.[CrossRef]
    [Google Scholar]
  8. Cashel, M., Gentry, D., Hernandez, V. J. & Vinella, D. ( 1996; ). The stringent response. In Escherichia coli and Salmonella: Cellular and Molecular Biology, pp. 1458–1496. Edited by F. C. Neidhardt, R. Curtiss III, J. L. Ingraham, E. C. C. Lin, K. B. Low, B. Magasanik, W. S. Reznikoff, M. Riley, M. Schaechter & H. E. Umbarger. Washington, DC: American Society for Microbiology.
  9. Cooper, T. F., Rozen, D. E. & Lenski, R. E. ( 2003; ). Parallel changes in gene expression after 20,000 generations of evolution in Escherichia coli. Proc Natl Acad Sci U S A 100, 1072–1077.[CrossRef]
    [Google Scholar]
  10. Ferenci, T. ( 2003; ). What is driving the acquisition of mutS and rpoS polymorphisms in Escherichia coli? Trends Microbiol 11, 457–461.[CrossRef]
    [Google Scholar]
  11. Ferenci, T. ( 2005; ). Maintaining a healthy SPANC balance through regulatory and mutational adaptation. Mol Microbiol 57, 1–8.[CrossRef]
    [Google Scholar]
  12. Gentry, D. R. & Cashel, M. ( 1996; ). Mutational analysis of the Escherichia coli spoT gene identifies distinct but overlapping regions involved in ppGpp synthesis and degradation. Mol Microbiol 19, 1373–1384.[CrossRef]
    [Google Scholar]
  13. Gentry, D. R., Hernadez, V. J., Nguyen, L. H., Jensen, D. B. & Cashel, M. ( 1993; ). Synthesis of stationary-phase sigma factor sigma-S is positively regulated by ppGpp. J Bacteriol 175, 7982–7989.
    [Google Scholar]
  14. Hengge-Aronis, R. ( 2002; ). Signal transduction and regulatory mechanisms involved in control of the sigma(S) (RpoS) subunit of RNA polymerase. Microbiol Mol Biol Rev 66, 373–395.[CrossRef]
    [Google Scholar]
  15. Hengge-Aronis, R. & Fischer, D. ( 1992; ). Identification and molecular analysis of glgS, a novel growth-phase-regulated and rpoS-dependent gene involved in glycogen synthesis in Escherichia coli. Mol Microbiol 6, 1877–1886.[CrossRef]
    [Google Scholar]
  16. Hirsch, M. & Elliott, T. ( 2002; ). Role of ppGpp in rpoS stationary-phase regulation in Escherichia coli. J Bacteriol 184, 5077–5087.[CrossRef]
    [Google Scholar]
  17. Hogg, T., Mechold, U., Malke, H., Cashel, M. & Hilgenfeld, R. ( 2004; ). Conformational antagonism between opposing active sites in a bifunctional RelA/SpoT homolog modulates (p)ppGpp metabolism during the stringent response. Cell 117, 57–68.[CrossRef]
    [Google Scholar]
  18. Jishage, M. & Ishihama, A. ( 1997; ). Variation in RNA polymerase sigma subunit composition within different stocks of Escherichia coli W3110. J Bacteriol 179, 959–963.
    [Google Scholar]
  19. Jishage, M., Kvint, K., Shingler, V. & Nystrom, T. ( 2002; ). Regulation of sigma factor competition by the alarmone ppGpp. Genes Dev 16, 1260–1270.[CrossRef]
    [Google Scholar]
  20. King, T., Ishihama, A., Kori, A. & Ferenci, T. ( 2004; ). A regulatory trade-off as a source of strain variation in the species Escherichia coli. J Bacteriol 186, 5614–5620.[CrossRef]
    [Google Scholar]
  21. King, T., Seeto, S. & Ferenci, T. ( 2006; ). Genotype-by-environment interactions influencing the emergence of rpoS mutations in Eschetichia coli populations. Genetics 172, 2071–2079.
    [Google Scholar]
  22. Kvint, K., Farewell, A. & Nystrom, T. ( 2000; ). RpoS-dependent promoters require guanosine tetraphosphate for induction even in the presence of high levels of sigma(s). J Biol Chem 275, 14795–14798.[CrossRef]
    [Google Scholar]
  23. Laffler, T. & Gallant, J. ( 1974; ). spoT, a new genetic locus involved in stringent response in E. coli. Cell 1, 27–30.[CrossRef]
    [Google Scholar]
  24. Lagosky, P. A. & Chang, F. N. ( 1980; ). Influence of amino-acid starvation on guanosine 5′-diphosphate 3′-diphosphate basal-level synthesis in Escherichia coli. J Bacteriol 144, 499–508.
    [Google Scholar]
  25. Lazzarini, R. A., Cashel, M. & Gallant, J. ( 1971; ). On the regulation of guanosine tetraphosphate levels in stringent and relaxed strains of Escherichia coli. J Biol Chem 246, 4381–4385.
    [Google Scholar]
  26. Magnusson, L. U., Farewell, A. & Nystrom, T. ( 2005; ). ppGpp: a global regulator in Escherichia coli. Trends Microbiol 13, 236–242.[CrossRef]
    [Google Scholar]
  27. Miller, J. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  28. Murray, K. D. & Bremer, H. ( 1996; ). Control of SpoT-dependent ppGpp synthesis and degradation in Escherichia coli. J Mol Biol 259, 41–57.[CrossRef]
    [Google Scholar]
  29. Nakanishi, N., Abe, H., Ogura, Y., Hayashi, T., Tashiro, K., Kuhara, S., Sugimoto, N. & Tobe, T. ( 2006; ). ppGpp with DksA controls gene expression in the locus of enterocyte effacement (LEE) pathogenicity island of enterohaemorrhagic Escherichia coli through activation of two virulence regulatory genes. Mol Microbiol 61, 194–205.[CrossRef]
    [Google Scholar]
  30. Notley-McRobb, L., King, T. & Ferenci, T. ( 2002; ). rpoS mutations and loss of general stress resistance in Escherichia coli populations as a consequence of conflict between competing stress responses. J Bacteriol 184, 806–811.[CrossRef]
    [Google Scholar]
  31. Nystrom, T. ( 2004; ). Growth versus maintenance: a trade-off dictated by RNA polymerase availability and sigma factor competition? Mol Microbiol 54, 855–862.[CrossRef]
    [Google Scholar]
  32. Reddy, P. S., Raghavan, A. & Chatterji, D. ( 1995; ). Evidence for a ppGpp-binding site on Escherichia coli RNA polymerase: proximity relationship with the rifampicin-binding domain. Mol Microbiol 15, 255–265.[CrossRef]
    [Google Scholar]
  33. Sarubbi, E., Rudd, K. E. & Cashel, M. ( 1988; ). Basal ppGpp level adjustment shown by new spoT mutants affect steady state growth rates and rrnA ribosomal promoter regulation in Escherichia coli. Mol Gen Genet 213, 214–222.[CrossRef]
    [Google Scholar]
  34. Sarubbi, E., Rudd, K. E., Xiao, H., Ikehara, K., Kalman, M. & Cashel, M. ( 1989; ). Characterization of the spoT gene of Escherichia coli. J Biol Chem 264, 15074–15082.
    [Google Scholar]
  35. Soupene, E., van Heeswijk, W. C., Plumbridge, J., Stewart, V., Bertenthal, D., Lee, H., Prasad, G., Paliy, O., Charernnoppakul, P. & Kustu, S. ( 2003; ). Physiological studies of Escherichia coli strain MG1655: growth defects and apparent cross-regulation of gene expression. J Bacteriol 185, 5611–5626.[CrossRef]
    [Google Scholar]
  36. Spira, B. & Ferenci, T. ( 2008; ). Alkaline phosphatase as a reporter of σ S levels and rpoS polymorphisms in different E. coli strains. Arch Microbiol 189, 43–47.
    [Google Scholar]
  37. Spira, B., Silberstein, N. & Yagil, E. ( 1995; ). Guanosine 3′,5′-bispyrophosphate (ppGpp) synthesis in cells of Escherichia coli starved for Pi. J Bacteriol 177, 4053–4058.
    [Google Scholar]
  38. Sutton, A., Buencamino, R. & Eisenstark, A. ( 2000; ). rpoS mutants in archival cultures of Salmonella enterica serovar Typhimurium. J Bacteriol 182, 4375–4379.[CrossRef]
    [Google Scholar]
  39. Typas, A., Becker, G. & Hengge, R. ( 2007; ). The molecular basis of selective promoter activation by the sigma(S) subunit of RNA polymerase. Mol Microbiol 63, 1296–1306.[CrossRef]
    [Google Scholar]
  40. Weber, H., Polen, T., Heuveling, J., Wendisch, V. F. & Hengge, R. ( 2005; ). Genome-wide analysis of the general stress response network in Escherichia coli: sigma(S)-dependent genes, promoters, and sigma factor selectivity. J Bacteriol 187, 1591–1603.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2008/018457-0
Loading
/content/journal/micro/10.1099/mic.0.2008/018457-0
Loading

Data & Media loading...

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