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. 1996 Mar;178(5):1394–1400. doi: 10.1128/jb.178.5.1394-1400.1996

Inorganic polyphosphate supports resistance and survival of stationary-phase Escherichia coli.

N N Rao 1, A Kornberg 1
PMCID: PMC177814  PMID: 8631717

Abstract

The Escherichia coli mutant (ppk) lacking the enzyme polyphosphate kinase, which makes long chains of inorganic polyphosphate (poly P), is deficient in functions expressed in the stationary phase of growth. After 2 days of growth in a medium limited in carbon sources, only 7% of the mutants survived compared with nearly 100% of the wild type; the loss in viability of the mutant was even more pronounced in a rich medium. The mutant showed a greater sensitivity to heat, to an oxidant (H2O2), to a redox-cycling agent (menadione), and to an osmotic challenge with 2.5 M NaCl. After a week or so in the stationary phase, mutant survivors were far fewer in number and were replaced by an outgrowth of a small-colony-size variant with a stable genotype and with improved viability and resistance to heat and H2O2; neither polyphosphate kinase nor long-chain poly P was restored. Suppression of the ppk feature of heat sensitivity by extra copies of rpoS, the gene encoding the RNA polymerase sigma factor that regulates some 50 stationary-phase genes, further implicates poly P in promoting survival in the stationary phase.

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Selected References

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  1. Akiyama M., Crooke E., Kornberg A. An exopolyphosphatase of Escherichia coli. The enzyme and its ppx gene in a polyphosphate operon. J Biol Chem. 1993 Jan 5;268(1):633–639. [PubMed] [Google Scholar]
  2. Akiyama M., Crooke E., Kornberg A. The polyphosphate kinase gene of Escherichia coli. Isolation and sequence of the ppk gene and membrane location of the protein. J Biol Chem. 1992 Nov 5;267(31):22556–22561. [PubMed] [Google Scholar]
  3. Almirón M., Link A. J., Furlong D., Kolter R. A novel DNA-binding protein with regulatory and protective roles in starved Escherichia coli. Genes Dev. 1992 Dec;6(12B):2646–2654. doi: 10.1101/gad.6.12b.2646. [DOI] [PubMed] [Google Scholar]
  4. Archibald F. S., Fridovich I. Investigations of the state of the manganese in Lactobacillus plantarum. Arch Biochem Biophys. 1982 May;215(2):589–596. doi: 10.1016/0003-9861(82)90120-5. [DOI] [PubMed] [Google Scholar]
  5. BEERS R. F., Jr, SIZER I. W. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952 Mar;195(1):133–140. [PubMed] [Google Scholar]
  6. Bohannon D. E., Connell N., Keener J., Tormo A., Espinosa-Urgel M., Zambrano M. M., Kolter R. Stationary-phase-inducible "gearbox" promoters: differential effects of katF mutations and role of sigma 70. J Bacteriol. 1991 Jul;173(14):4482–4492. doi: 10.1128/jb.173.14.4482-4492.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bol D. K., Yasbin R. E. Characterization of an inducible oxidative stress system in Bacillus subtilis. J Bacteriol. 1990 Jun;172(6):3503–3506. doi: 10.1128/jb.172.6.3503-3506.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bonting C. F., Kortstee G. J., Zehnder A. J. Properties of polyphosphate: AMP phosphotransferase of Acinetobacter strain 210A. J Bacteriol. 1991 Oct;173(20):6484–6488. doi: 10.1128/jb.173.20.6484-6488.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clare D. A., Duong M. N., Darr D., Archibald F., Fridovich I. Effects of molecular oxygen on detection of superoxide radical with nitroblue tetrazolium and on activity stains for catalase. Anal Biochem. 1984 Aug 1;140(2):532–537. doi: 10.1016/0003-2697(84)90204-5. [DOI] [PubMed] [Google Scholar]
  10. Crooke E., Akiyama M., Rao N. N., Kornberg A. Genetically altered levels of inorganic polyphosphate in Escherichia coli. J Biol Chem. 1994 Mar 4;269(9):6290–6295. [PubMed] [Google Scholar]
  11. DIRHEIMER G., EBEL J. P. SUR LE M'ETABOLISME DES POLYPHOSPHATES INORGANIQUES CHEZ CORYNEBACTERIUM XEROSIS. C R Seances Soc Biol Fil. 1964;158:1948–1951. [PubMed] [Google Scholar]
  12. Demple B., Halbrook J. Inducible repair of oxidative DNA damage in Escherichia coli. Nature. 1983 Aug 4;304(5925):466–468. doi: 10.1038/304466a0. [DOI] [PubMed] [Google Scholar]
  13. Demple B. Regulation of bacterial oxidative stress genes. Annu Rev Genet. 1991;25:315–337. doi: 10.1146/annurev.ge.25.120191.001531. [DOI] [PubMed] [Google Scholar]
  14. Dunn T., Gable K., Beeler T. Regulation of cellular Ca2+ by yeast vacuoles. J Biol Chem. 1994 Mar 11;269(10):7273–7278. [PubMed] [Google Scholar]
  15. Fridovich I. The biology of oxygen radicals. Science. 1978 Sep 8;201(4359):875–880. doi: 10.1126/science.210504. [DOI] [PubMed] [Google Scholar]
  16. Gentry D. R., Hernandez V. J., Nguyen L. H., Jensen D. B., Cashel M. Synthesis of the stationary-phase sigma factor sigma s is positively regulated by ppGpp. J Bacteriol. 1993 Dec;175(24):7982–7989. doi: 10.1128/jb.175.24.7982-7989.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Greenberg J. T., Demple B. A global response induced in Escherichia coli by redox-cycling agents overlaps with that induced by peroxide stress. J Bacteriol. 1989 Jul;171(7):3933–3939. doi: 10.1128/jb.171.7.3933-3939.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Greenberg J. T., Demple B. Overproduction of peroxide-scavenging enzymes in Escherichia coli suppresses spontaneous mutagenesis and sensitivity to redox-cycling agents in oxyR-mutants. EMBO J. 1988 Aug;7(8):2611–2617. doi: 10.1002/j.1460-2075.1988.tb03111.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hengge-Aronis R., Klein W., Lange R., Rimmele M., Boos W. Trehalose synthesis genes are controlled by the putative sigma factor encoded by rpoS and are involved in stationary-phase thermotolerance in Escherichia coli. J Bacteriol. 1991 Dec;173(24):7918–7924. doi: 10.1128/jb.173.24.7918-7924.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hengge-Aronis R., Lange R., Henneberg N., Fischer D. Osmotic regulation of rpoS-dependent genes in Escherichia coli. J Bacteriol. 1993 Jan;175(1):259–265. doi: 10.1128/jb.175.1.259-265.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hengge-Aronis R. Survival of hunger and stress: the role of rpoS in early stationary phase gene regulation in E. coli. Cell. 1993 Jan 29;72(2):165–168. doi: 10.1016/0092-8674(93)90655-a. [DOI] [PubMed] [Google Scholar]
  22. Hsieh P. C., Shenoy B. C., Jentoft J. E., Phillips N. F. Purification of polyphosphate and ATP glucose phosphotransferase from Mycobacterium tuberculosis H37Ra: evidence that poly(P) and ATP glucokinase activities are catalyzed by the same enzyme. Protein Expr Purif. 1993 Feb;4(1):76–84. doi: 10.1006/prep.1993.1012. [DOI] [PubMed] [Google Scholar]
  23. Imlay J. A., Chin S. M., Linn S. Toxic DNA damage by hydrogen peroxide through the Fenton reaction in vivo and in vitro. Science. 1988 Apr 29;240(4852):640–642. doi: 10.1126/science.2834821. [DOI] [PubMed] [Google Scholar]
  24. Imlay J. A., Linn S. DNA damage and oxygen radical toxicity. Science. 1988 Jun 3;240(4857):1302–1309. doi: 10.1126/science.3287616. [DOI] [PubMed] [Google Scholar]
  25. Ivanova A., Miller C., Glinsky G., Eisenstark A. Role of rpoS (katF) in oxyR-independent regulation of hydroperoxidase I in Escherichia coli. Mol Microbiol. 1994 May;12(4):571–578. doi: 10.1111/j.1365-2958.1994.tb01043.x. [DOI] [PubMed] [Google Scholar]
  26. Jenkins D. E., Chaisson S. A., Matin A. Starvation-induced cross protection against osmotic challenge in Escherichia coli. J Bacteriol. 1990 May;172(5):2779–2781. doi: 10.1128/jb.172.5.2779-2781.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jenkins D. E., Schultz J. E., Matin A. Starvation-induced cross protection against heat or H2O2 challenge in Escherichia coli. J Bacteriol. 1988 Sep;170(9):3910–3914. doi: 10.1128/jb.170.9.3910-3914.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Keasling J. D., Bertsch L., Kornberg A. Guanosine pentaphosphate phosphohydrolase of Escherichia coli is a long-chain exopolyphosphatase. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7029–7033. doi: 10.1073/pnas.90.15.7029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kornberg A. Inorganic polyphosphate: toward making a forgotten polymer unforgettable. J Bacteriol. 1995 Feb;177(3):491–496. doi: 10.1128/jb.177.3.491-496.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lange R., Barth M., Hengge-Aronis R. Complex transcriptional control of the sigma s-dependent stationary-phase-induced and osmotically regulated osmY (csi-5) gene suggests novel roles for Lrp, cyclic AMP (cAMP) receptor protein-cAMP complex, and integration host factor in the stationary-phase response of Escherichia coli. J Bacteriol. 1993 Dec;175(24):7910–7917. doi: 10.1128/jb.175.24.7910-7917.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lange R., Hengge-Aronis R. Growth phase-regulated expression of bolA and morphology of stationary-phase Escherichia coli cells are controlled by the novel sigma factor sigma S. J Bacteriol. 1991 Jul;173(14):4474–4481. doi: 10.1128/jb.173.14.4474-4481.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Li C., Clarke S. A protein methyltransferase specific for altered aspartyl residues is important in Escherichia coli stationary-phase survival and heat-shock resistance. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9885–9889. doi: 10.1073/pnas.89.20.9885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Loewen P. C., Switala J., Triggs-Raine B. L. Catalases HPI and HPII in Escherichia coli are induced independently. Arch Biochem Biophys. 1985 Nov 15;243(1):144–149. doi: 10.1016/0003-9861(85)90782-9. [DOI] [PubMed] [Google Scholar]
  34. McCann M. P., Kidwell J. P., Matin A. The putative sigma factor KatF has a central role in development of starvation-mediated general resistance in Escherichia coli. J Bacteriol. 1991 Jul;173(13):4188–4194. doi: 10.1128/jb.173.13.4188-4194.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Mulvey M. R., Sorby P. A., Triggs-Raine B. L., Loewen P. C. Cloning and physical characterization of katE and katF required for catalase HPII expression in Escherichia coli. Gene. 1988 Dec 20;73(2):337–345. doi: 10.1016/0378-1119(88)90498-2. [DOI] [PubMed] [Google Scholar]
  36. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Phillips N. F., Horn P. J., Wood H. G. The polyphosphate- and ATP-dependent glucokinase from Propionibacterium shermanii: both activities are catalyzed by the same protein. Arch Biochem Biophys. 1993 Jan;300(1):309–319. doi: 10.1006/abbi.1993.1043. [DOI] [PubMed] [Google Scholar]
  38. Pick U., Bental M., Chitlaru E., Weiss M. Polyphosphate-hydrolysis--a protective mechanism against alkaline stress? FEBS Lett. 1990 Nov 12;274(1-2):15–18. doi: 10.1016/0014-5793(90)81318-i. [DOI] [PubMed] [Google Scholar]
  39. Pick U., Weiss M. Polyphosphate Hydrolysis within Acidic Vacuoles in Response to Amine-Induced Alkaline Stress in the Halotolerant Alga Dunaliella salina. Plant Physiol. 1991 Nov;97(3):1234–1240. doi: 10.1104/pp.97.3.1234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Rao N. N., Roberts M. F., Torriani A. Amount and chain length of polyphosphates in Escherichia coli depend on cell growth conditions. J Bacteriol. 1985 Apr;162(1):242–247. doi: 10.1128/jb.162.1.242-247.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Reusch R. N., Sadoff H. L. Putative structure and functions of a poly-beta-hydroxybutyrate/calcium polyphosphate channel in bacterial plasma membranes. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4176–4180. doi: 10.1073/pnas.85.12.4176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Roszak D. B., Colwell R. R. Survival strategies of bacteria in the natural environment. Microbiol Rev. 1987 Sep;51(3):365–379. doi: 10.1128/mr.51.3.365-379.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Siegele D. A., Kolter R. Isolation and characterization of an Escherichia coli mutant defective in resuming growth after starvation. Genes Dev. 1993 Dec;7(12B):2629–2640. doi: 10.1101/gad.7.12b.2629. [DOI] [PubMed] [Google Scholar]
  44. Siegele D. A., Kolter R. Life after log. J Bacteriol. 1992 Jan;174(2):345–348. doi: 10.1128/jb.174.2.345-348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tanaka K., Takayanagi Y., Fujita N., Ishihama A., Takahashi H. Heterogeneity of the principal sigma factor in Escherichia coli: the rpoS gene product, sigma 38, is a second principal sigma factor of RNA polymerase in stationary-phase Escherichia coli. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3511–3515. doi: 10.1073/pnas.90.8.3511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Thor H., Smith M. T., Hartzell P., Bellomo G., Jewell S. A., Orrenius S. The metabolism of menadione (2-methyl-1,4-naphthoquinone) by isolated hepatocytes. A study of the implications of oxidative stress in intact cells. J Biol Chem. 1982 Oct 25;257(20):12419–12425. [PubMed] [Google Scholar]
  47. Tormo A., Almirón M., Kolter R. surA, an Escherichia coli gene essential for survival in stationary phase. J Bacteriol. 1990 Aug;172(8):4339–4347. doi: 10.1128/jb.172.8.4339-4347.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Van Veen H. W., Abee T., Kortstee G. J., Konings W. N., Zehnder A. J. Characterization of two phosphate transport systems in Acinetobacter johnsonii 210A. J Bacteriol. 1993 Jan;175(1):200–206. doi: 10.1128/jb.175.1.200-206.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wada A., Yamazaki Y., Fujita N., Ishihama A. Structure and probable genetic location of a "ribosome modulation factor" associated with 100S ribosomes in stationary-phase Escherichia coli cells. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2657–2661. doi: 10.1073/pnas.87.7.2657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Weichart D., Lange R., Henneberg N., Hengge-Aronis R. Identification and characterization of stationary phase-inducible genes in Escherichia coli. Mol Microbiol. 1993 Oct;10(2):407–420. [PubMed] [Google Scholar]
  51. Wood H. G., Clark J. E. Biological aspects of inorganic polyphosphates. Annu Rev Biochem. 1988;57:235–260. doi: 10.1146/annurev.bi.57.070188.001315. [DOI] [PubMed] [Google Scholar]
  52. Wu J., Weiss B. Two divergently transcribed genes, soxR and soxS, control a superoxide response regulon of Escherichia coli. J Bacteriol. 1991 May;173(9):2864–2871. doi: 10.1128/jb.173.9.2864-2871.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wurst H., Shiba T., Kornberg A. The gene for a major exopolyphosphatase of Saccharomyces cerevisiae. J Bacteriol. 1995 Feb;177(4):898–906. doi: 10.1128/jb.177.4.898-906.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Yamagishi M., Matsushima H., Wada A., Sakagami M., Fujita N., Ishihama A. Regulation of the Escherichia coli rmf gene encoding the ribosome modulation factor: growth phase- and growth rate-dependent control. EMBO J. 1993 Feb;12(2):625–630. doi: 10.1002/j.1460-2075.1993.tb05695.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Zambrano M. M., Siegele D. A., Almirón M., Tormo A., Kolter R. Microbial competition: Escherichia coli mutants that take over stationary phase cultures. Science. 1993 Mar 19;259(5102):1757–1760. doi: 10.1126/science.7681219. [DOI] [PubMed] [Google Scholar]
  56. van Groenestijn J. W., Zuidema M., van de Worp J. J., Deinema M. H., Zehnder A. J. Influence of environmental parameters on polyphosphate accumulation in Acinetobacter sp. Antonie Van Leeuwenhoek. 1989;55(1):67–82. doi: 10.1007/BF02309620. [DOI] [PubMed] [Google Scholar]

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