Legionella pneumophila CsrA is a pivotal repressor of transmission traits and activator of replication

Abstract

Summary Legionella pneumophila can replicate inside amoebae and also alveolar macrophages to cause Legionnaires’ Disease in susceptible hosts. When nutrients become limiting, a stringent‐like response coordinates the differentiation of L. pneumophila to a transmissive form, a process mediated by the two‐component system LetA/S and the sigma factors RpoS and FliA. Here we demonstrate that the broadly conserved RNA binding protein CsrA is a global repressor of L. pneumophila transmission phenotypes and an essential activator of intracellular replication. By analysing csrA expression and the phenotypes of csrA single and double mutants and a strain that expresses csrA constitutively, we demonstrate that, during replication in broth, CsrA represses every post‐exponential phase phenotype examined, including cell shape shortening, motility, pigmentation, stress resistance, sodium sensitivity, cytotoxicity and efficient macrophage infection. At the transition to the post‐exponential phase, LetA/S relieves CsrA repression to induce transmission phenotypes by both FliA‐dependent and ‐independent pathways. For L. pneumophila to avoid lysosomal degradation in macrophages, CsrA repression must be relieved by LetA/S before phagocytosis; conversely, before intracellular bacteria can replicate, CsrA repression must be restored. The reciprocal regulation of replication and transmission exemplified by CsrA likely enhances the fitness of microbes faced with fluctuating environments.

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References 69

1. 1.

   Aarons S, Abbas A, Adams C, Fenton A, and O’Gara F (2000 ) A regulatory RNA (PrrB RNA) modulates expression of secondary metabolite genes in Pseudomonas fluorescens F113. J Bacteriol 182: 3913–3919.

   Not in our library10.1128/jb.182.14.3913-3919.2000

2. 2.

   Alli OAT, Gao L-Y, Pedersen LL, Zink S, Radulic M, Doric M, and Abu Kwaik Y (2000 ) Temporal pore formation-mediated egress from macrophages and alveolar epithelial cells by Legionella pneumophila. Infect Immun 68: 6431–6440.

   Not in our library10.1128/iai.68.11.6431-6440.2000

3. 3.

   Altier C, Suyemoto M, and Lawhon SD (2000a ) Regulation of Salmonella enterica serovar typhimurium invasion genes by csrA. Infect Immun 68: 6790–6797.

   Not in our library10.1128/iai.68.12.6790-6797.2000

4. 4.

   Altier C, Suyemoto M, Ruiz AI, Burnham KD, and Maurer R (2000b ) Characterization of two novel regulatory genes affecting Salmonella invasion gene expression. Mol Microbiol 35: 635–646.

   Not in our library10.1046/j.1365-2958.2000.01734.x

5. 5.

   Bachman MA, and Swanson MS (2001 ) RpoS cooperates with other factors to induce Legionella pneumophila virulence in the stationary phase. Mol Microbiol 40: 1201–1214.

   Not in our library10.1046/j.1365-2958.2001.02465.x

6. 6.

   Baker CS, Morozov I, Suzuki K, Romeo T, and Babitzke P (2002 ) CsrA regulates glycogen biosynthesis by preventing translation of glgC in Escherichia coli. Mol Microbiol 44: 1599–1610.

   Not in our library10.1046/j.1365-2958.2002.02982.x

7. 7.

   Bandyopadhyay P, and Steinman HM (1998 ) Legionella pneumophila catalase-peroxidases: cloning of the katB gene and studies of KatB function. J Bacteriol 180: 5369–5374.

   Not in our library10.1128/jb.180.20.5369-5374.1998

8. 8.

   Berger KH, and Isberg RR (1993 ) Two distinct defects in intracellular growth complemented by a single genetic locus in Legionella pneumophila. Mol Micro 7: 7–19.

   Not in our library10.1111/j.1365-2958.1993.tb01092.x

9. 9.

   Berger KH, Merriam JJ, and Isberg RI (1994 ) Altered intracellular targeting properties associated with mutations in the Legionella pneumophila dotA gene. Mol Microbiol 14: 809–822.

   Not in our library10.1111/j.1365-2958.1994.tb01317.x

10. 10.

    Blumer C, Heeb S, Pessi G, and Haas D (1999 ) Global GacA-steered control of cyanide and exoprotease production in Pseudomonas fluorescens involves specific ribosome binding sites. Proc Natl Acad Sci USA 96: 14073–14078.

    Not in our library10.1073/pnas.96.24.14073

11. 11.

    Byrne B, and Swanson MS (1998 ) Expression of Legionella pneumophila virulence traits in response to growth conditions. Infect Immun 66: 3029–3034.

    Not in our library10.1128/iai.66.7.3029-3034.1998

12. 12.

    Catrenich CE, and Johnson W (1989 ) Characterization of the selective inhibition of growth of virulent Legionella pneumophila by supplemented Mueller-Hinton medium. Infect Immun 57: 1862–1864.

    Not in our library10.1128/iai.57.6.1862-1864.1989

13. 13.

    Chatterjee A, Cui Y, Liu Y, Dumenyo CK, and Chatterjee AK (1995 ) Inactivation of rsmA leads to overproduction of extracellular pectinases, cellulases, and proteases in Erwinia carotovora subsp. carotovora in the absence of the starvation/cell density-sensing signal, N-(3-oxohexanoyl)-l-homoserine lactone. Appl Environ Microbiol 61: 1959–1967.

    Not in our library10.1128/aem.61.5.1959-1967.1995

14. 14.

    Chilcott GS, and Hughes KT (2000 ) Coupling of flagellar gene expression to flagellar assembly in Salmonella enterica serovar typhimurium and Escherichia coli. Microbiol Mol Biol Rev 64: 694–708.

    Not in our library10.1128/mmbr.64.4.694-708.2000

15. 15.

    Coers J, Monahan C, and Roy CR (1999 ) Modulation of phagosome biogenesis by Legionella pneumophila creates an organelle permissive for intracellular growth. Nature Cell Biol 7: 451–453.

    Not in our library10.1038/15687

16. 16.

    Cui Y, Chatterjee A, Liu Y, Dumenyo CK, and Chatterjee AK (1995 ) Identification of a global repressor gene, rsmA, of Erwinia carotovora subsp. carotovora that controls extracellular enzymes, N-(3-oxohexanoyl)-l-homoserine lactone, and pathogenicity in soft-rotting Erwinia spp. J Bacteriol 177: 5108–5115.

    Not in our library10.1128/jb.177.17.5108-5115.1995

17. 17.

    Cui Y, Chatterjee A, and Chatterjee AK (2001 ) Effects of the two-component system comprising GacA and GacS of Erwinia carotovora subsp. carotovora on the production of global regulatory rsmB RNA, extracellular enzymes, and harpinEcc. Mol Plant Microbe Interact 14: 516–526.

    Not in our library10.1094/mpmi.2001.14.4.516

18. 18.

    Eichelberg K, and Galan JE (2000 ) The flagellar sigma factor FliA (sigma (28) regulates the expression of Salmonella genes associated with the centisome 63 type III secretion system. Infect Immun 68: 2735–2743.

    Not in our library10.1128/iai.68.5.2735-2743.2000

19. 19.

    Faulkner G, and Garduno RA (2002 ) Ultrastructural analysis of differentiation in Legionella pneumophila. J Bacteriol 184: 7025–7041.

    Not in our library10.1128/jb.184.24.7025-7041.2002

20. 20.

    Fettes PS, Forsbach-Birk V, Lynch D, and Marre R (2001 ) Overexpresssion of a Legionella pneumophila homologue of the E. coli regulator csrA affects cell size, flagellation, and pigmentation. Int J Med Microbiol 291: 353–360.

    Not in our library10.1078/1438-4221-00141

21. 21.

    Fields BS (1996 ) The molecular ecology of legionellae. Trends Micro 4: 286–290.

    Not in our library10.1016/0966-842x(96)10041-x

22. 22.

    Frey J, Bagdasarian M, Feiss D, Frankline FCH, and Deshusses J (1983 ) Stable cosmid vectors that enable the introduction of cloned fragments into a wide range of Gram-negative bacteria. Gene 24: 299–308.

    Not in our library10.1016/0378-1119(83)90090-2

23. 23.

    Garduno RA, Garduno E, Hiltz M, and Hoffman PS (2002 ) Intracellular growth of Legionella pneumophila gives rise to a differentiated form dissimilar to stationary-phase forms. Infect Immun 70: 6273–6283.

    Not in our library10.1128/iai.70.11.6273-6283.2002

24. 24.

    George JR, Pine L, Reeves MW, and Harrell WK (1980 ) Amino acid requirements of Legionella pneumophila. J Clin Microbiol 11: 286–291.

    Not in our library10.1128/jcm.11.3.286-291.1980

25. 25.

    Hales LM, and Shuman HA (1999 ) The Legionella pneumophila rpoS gene is required for growth within Acanthamoeba castellanii. J Bacteriol 181: 4879–4889.

    Not in our library10.1128/jb.181.16.4879-4889.1999

26. 26.

    Hammer BK, and Swanson MS (1999 ) Co-ordination of Legionella pneumophila virulence with entry into stationary phase by ppGpp. Mol Microbiol 33: 721–731.

    Not in our library10.1046/j.1365-2958.1999.01519.x

27. 27.

    Hammer BK, Suzuki E, and Swanson M (2002 ) A two-component regulator induces the transmission phenotype of stationary phase Legionella pneumophila. Mol Microbiol 44: 107–118.

    Not in our library10.1046/j.1365-2958.2002.02884.x

28. 28.

    Heeb S, and Haas D (2001 ) Regulatory roles of the GacS/GacA two-component system in plant-associated and other Gram-negative bacteria. Mol Plant Microbe Interact 14: 1351–1363.

    Not in our library10.1094/mpmi.2001.14.12.1351

29. 29.

    Heeb S, Blumer C, and Haas D (2002 ) Regulatory RNA as mediator in GacA/RsmA-dependent global control of exoproduct formation in Pseudomonas fluorescens CHA0. J Bacteriol 184: 1046–1056.

    Not in our library10.1128/jb.184.4.1046-1056.2002

30. 30.

    Heuner K, Bender-Beck L, Brand BC, Luck PC, Mann KH, Marre R, (1995 ) Cloning and genetic characterization of the flagellum subunit gene (flaA) of Legionella pneumophila serogroup 1. Infect Immun 63: 2499–2507.

    Not in our library10.1128/iai.63.7.2499-2507.1995

31. 31.

    Heuner K, Hacker J, and Brand BC (1997 ) The alternative sigma factor sigma28 of Legionella pneumophila restores flagellation and motility to an Escherichia coli fliA mutant. J Bacteriol 179: 17–23.

    Not in our library10.1128/jb.179.1.17-23.1997

32. 32.

    Heuner K, Brand BC, and Hacker J (1999 ) The expression of the flagellum of Legionella pneumophila is modulated by different environmental factors. FEMS Microbiol Lett 175: 69–77.

    Not in our library10.1111/j.1574-6968.1999.tb13603.x

33. 33.

    Heuner K, Dietrich C, Skriwan C, Steinert M, and Hacker J (2002 ) Influence of the alternative sigma (28) factor on virulence and flagellum expression of Legionella pneumophila. Infect Immun 70: 1604–1608.

    Not in our library10.1128/iai.70.3.1604-1608.2002

34. 34.

    Hoffman P (1984 ) Bacterial physiology. In Legionella: Proceedings of the 2nd International Symposium. Thornsberry C, Balows A, Feeley JC, and Jakubowski W, (eds). Washington, D.C.: American Society for Microbiology, pp. 61–67.

    Not in our library

35. 35.

    Horwitz MA (1983 ) The Legionnaires’ disease bacterium (Legionella pneumophila) inhibits phagosome lysosome fusion in human monocytes. J Exp Med 158: 2108–2126.

    Not in our library10.1084/jem.158.6.2108

36. 36.

    Horwitz MA, and Silverstein SC (1980 ) Legionnaires’ disease bacterium (Legionella pneumophila) multiplies intracellularly in human monocytes. J Clin Invest 66: 441–450.

    Not in our library10.1172/jci109874

37. 37.

    Jackson DW, Suzuki K, Oakford L, Simecka JW, Hart ME, and Romeo T (2002 ) Biofilm formation and dispersal under the influence of the global regulator CsrA of Escherichia coli. J Bacteriol 184: 290–301.

    Not in our library10.1128/jb.184.1.290-301.2002

38. 38.

    Kirby JE, Vogel JP, Andrews HL, and Isberg RR (1998 ) Evidence of pore-forming ability by Legionella pneumophila. Mol Microbiol 27: 323–336.

    Not in our library10.1046/j.1365-2958.1998.00680.x

39. 39.

    Lynch D, Fieser N, Gloggler K, Forsbach-Birk V, and Marre R (2003 ) The response regulator LetA regulates the stationary-phase stress response in Legionella pneumophila and is required for efficient infection of Acanthamoeba castellanii. FEMS Microbiol Lett 219: 241–248.

    Not in our library10.1016/s0378-1097(03)00050-8

40. 40.

    McDade JE, Shepard CC, Fraser DW, Tsai TR, Redus MA, and Dowdle WR (1977 ) Legionnaires’ disease: isolation of a bacterium and demonstration of its role in other respiratory diseases. N Engl J Med 297: 1197–1203.

    Not in our library10.1056/nejm197712012972202

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