Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Review Article
  • Published:

Life on the inside: the intracellular lifestyle of cytosolic bacteria

Nature Reviews Microbiology volume 7, pages 333–340 (2009)Cite this article

Key Points

  • Bacterial pathogens exploit a range of niches within their hosts. A small number of bacteria, including Listeria monocytogenes, Shigella flexneri, Burkholderia pseudomallei, Francisella tularensis and Rickettsia spp., are able to gain access to and proliferate in the cell cytosol and are termed cytosolic bacteria.

  • Escape from the vacuole following invasion is crucial for cytosolic pathogens. Bacteria escape rapidly from the vacuole through mechanisms that rely on the production of secreted enzymes and form pores to disrupt the vacuolar membrane. Interestingly, with the exception of F. tularensis, all cytosolic bacteria use actin-based motility after entry into the cytosol and spread to neighbouring cells.

  • Little is known about the biochemical composition of the mammalian cell cytosol and a key, but unresolved, question is whether the cytosol is permissive for bacterial growth.

  • Cytosolic bacteria are adapted to replicate in the cell cytosol. Studies identifying the bacterial genes and growth requirements that are important for intracellular replication have been informative regarding the nutrient availability within the cytosol. The cytosol seems to be limiting for compounds such as aromatic amino acids, and bacteria can readily use carbon sources, including pyruvate (by Rickettsia spp.) and hexose phosphates (by S. flexneri and L. monocytogenes), that must be available to microorganisms in the cytosol.

  • Bacteria in the cytosol are recognized by the innate immune system and autophagy is a key component of the host defence against cytosolic bacteria. There is increasing evidence that cytosolic bacteria interact and modify the autophagic pathway to promote their survival.

  • Cytosolic bacteria have evolved several mechanisms to adapt to their preferred niche. Future work on these pathogens will provide information on the cytosol as a site for replication and the bacterial strategies required to survive within it. Furthermore, it is becoming appreciated that a wider range of bacteria can exploit this host niche during steps in their pathogenesis.

Abstract

Bacterial pathogens exploit a huge range of niches within their hosts. Many pathogens can invade non-phagocytic cells and survive within a membrane-bound compartment. However, only a small number of bacteria, including Listeria monocytogenes, Shigella flexneri, Burkholderia pseudomallei, Francisella tularensis and Rickettsia spp., can gain access to and proliferate within the host cell cytosol. Here, we discuss the mechanisms by which these cytosolic pathogens escape into the cytosol, obtain nutrients to replicate and subvert host immune responses.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: The intracellular lifestyle of cytosolic pathogens.
Figure 2: Intracellular replication of bacteria within the cytosol of mammalian cells.
Figure 3: Interaction of cytosolic bacteria with the autophagic pathway.

Similar content being viewed by others

References

  1. Cossart, P. & Sansonetti, P. J. Bacterial invasion: the paradigms of enteroinvasive pathogens. Science 304, 242–248 (2004).

    Article  CAS  PubMed  Google Scholar 

  2. Haas, A. The phagosome: compartment with a license to kill. Traffic 8, 311–330 (2007).

    Article  CAS  PubMed  Google Scholar 

  3. Sansonetti, P. J. Rupture, invasion and inflammatory destruction of the intestinal barrier by Shigella, making sense of prokaryote–eukaryote cross-talks. FEMS Microbiol. Rev. 25, 3–14 (2001).

    CAS  PubMed  Google Scholar 

  4. Schroeder, G. N. & Hilbi, H. Molecular pathogenesis of Shigella spp.: controlling host cell signaling, invasion, and death by type III secretion. Clin. Microbiol. Rev. 21, 134–156 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Beauregard, K. E., Lee, K. D., Collier, R. J. & Swanson, J. A. pH-dependent perforation of macrophage phagosomes by listeriolysin O from Listeria monocytogenes. Mol. Biol. Cell 8, 680 (1997).

    Google Scholar 

  6. Golovliov, I., Baranov, V., Krocova, Z., Kovarova, H. & Sjostedt, A. An attenuated strain of the facultative intracellular bacterium Francisella tularensis can escape the phagosome of monocytic cells. Infect. Immun. 71, 5940–5950 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Henry, R. et al. Cytolysin-dependent delay of vacuole maturation in macrophages infected with Listeria monocytogenes. Cell. Microbiol. 8, 107–119 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Myers, J. T., Tsang, A. W. & Swanson, J. A. Localized reactive oxygen and nitrogen intermediates inhibit escape of Listeria monocytogenes from vacuoles in activated macrophages. J. Immunol. 171, 5447–5453 (2003).

    Article  CAS  PubMed  Google Scholar 

  9. Sansonetti, P. J., Ryter, A., Clerc, P., Maurelli, A. T. & Mounier, J. Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis. Infect. Immun. 51, 461–469 (1986). First study to show that S. flexneri escapes the vacuole and proliferates in the cytosol.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Santic, M., Asare, R., Skrobonja, I., Jones, S. & Abu Kwaik, Y. Acquisition of the vacuolar ATPase proton pump and phagosome acidification are essential for escape of Francisella tularensis into the macrophage cytosol. Infect. Immun. 76, 2671–2677 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Shaughnessy, L. M., Hoppe, A. D., Christensen, J. A. & Swanson, J. A. Membrane perforations inhibit lysosome fusion by altering pH and calcium in Listeria monocytogenes vacuoles. Cell. Microbiol. 8, 781–792 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Teysseire, N., Boudier, J. A. & Raoult, D. Rickettsia conorii entry into Vero cells. Infect. Immun. 63, 366–374 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Tilney, L. G. & Portnoy, D. A. Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes. J. Cell Biol. 109, 1597–1608 (1989).

    Article  CAS  PubMed  Google Scholar 

  14. Yates, R. M., Hermetter, A. & Russell, D. G. The kinetics of phagosome maturation as a function of phagosome/lysosome fusion and acquisition of hydrolytic activity. Traffic 6, 413–420 (2005).

    Article  CAS  PubMed  Google Scholar 

  15. Cossart, P. et al. Listeriolysin O is essential for virulence of Listeria monocytogenes: direct evidence obtained by gene complementation. Infect. Immun. 57, 3629–3636 (1989).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Gedde, M. M., Higgins, D. E., Tilney, L. G. & Portnoy, D. A. Role of listeriolysin O in cell-to-cell spread of Listeria monocytogenes. Infect. Immun. 68, 999–1003 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Portnoy, D. A., Jacks, P. S. & Hinrichs, D. J. Role of hemolysin for the intracellular growth of Listeria monocytogenes. J. Exp. Med. 167, 1459–1471 (1988).

    Article  CAS  PubMed  Google Scholar 

  18. Smith, G. A. et al. The two distinct phospholipases C of Listeria monocytogenes have overlapping roles in escape from a vacuole and cell-to-cell spread. Infect. Immun. 63, 4231–4237 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Bielecki, J., Youngman, P., Connelly, P. & Portnoy, D. A. Bacillus subtilis expressing a hemolysin gene from Listeria monocytogenes can grow in mammalian cells. Nature 345, 175–176 (1990). This key paper reported that expression of LLO is sufficient to allow vacuole escape and that the mammalian cell cytosol is permissive for bacterial growth.

    Article  CAS  PubMed  Google Scholar 

  20. Portnoy, D. A., Tweten, R. K., Kehoe, M. & Bielecki, J. Capacity of listeriolysin O, streptolysin O, and perfringolysin O to mediate growth of Bacillus subtilis within mammalian cells. Infect. Immun. 60, 2710–2717 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Camilli, A., Tilney, L. G. & Portnoy, D. A. Dual roles of plcA in Listeria monocytogenes pathogenesis. Mol. Microbiol. 8, 143–157 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Marquis, H. & Hager, E. J. pH-regulated activation and release of a bacteria-associated phospholipase C during intracellular infection by Listeria monocytogenes. Mol. Microbiol. 35, 289–298 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Schnupf, P. et al. Regulated translation of listeriolysin O controls virulence of Listeria monocytogenes. Mol. Microbiol. 61, 999–1012 (2006).

    Article  CAS  PubMed  Google Scholar 

  24. Glomski, I. J., Decatur, A. L. & Portnoy, D. A. Listeria monocytogenes mutants that fail to compartmentalize listerolysin O activity are cytotoxic, avirulent, and unable to evade host extracellular defenses. Infect. Immun. 71, 6754–6765 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Glomski, I. J., Gedde, M. M., Tsang, A. W., Swanson, J. A. & Portnoy, D. A. The Listeria monocytogenes hemolysin has an acidic pH optimum to compartmentalize activity and prevent damage to infected host cells. J. Cell Biol. 156, 1029–1038 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Singh, R., Jamieson, A. & Cresswell, P. GILT is a critical host factor for Listeria monocytogenes infection. Nature 455, 1244–1247 (2008). This study shows that LLO is activated by a host factor present in the vacuole.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Geoffroy, C., Gaillard, J. L., Alouf, J. E. & Berche, P. Purification, characterization, and toxicity of the sulfhydryl-activated hemolysin listeriolysin O from Listeria monocytogenes. Infect. Immun. 55, 1641–1646 (1987).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. High, N., Mounier, J., Prevost, M. C. & Sansonetti, P. J. IpaB of Shigella flexneri causes entry into epithelial cells and escape from the phagocytic vacuole. EMBO J. 11, 1991–1999 (1992).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Hayward, R. D. et al. Cholesterol binding by the bacterial type III translocon is essential for virulence effector delivery into mammalian cells. Mol. Microbiol. 56, 590–603 (2005).

    Article  CAS  PubMed  Google Scholar 

  30. Picking, W. L. et al. IpaD of Shigella flexneri is independently required for regulation of Ipa protein secretion and efficient insertion of IpaB and IpaC into host membranes. Infect. Immun. 73, 1432–1440 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Maurelli, A. T., Baudry, B., Dhauteville, H., Hale, T. L. & Sansonetti, P. J. Cloning of plasmid DNA sequences involved in invasion of HeLa cells by Shigella flexneri. Infect. Immun. 49, 164–171 (1985).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Sansonetti, P. J., Kopecko, D. J. & Formal, S. B. Involvement of a plasmid in the invasive ability of Shigella flexneri. Infect. Immun. 35, 852–860 (1982).

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Fernandez-Prada, C. M. et al. Shigella flexneri IpaH7.8 facilitates escape of virulent bacteria from the endocytic vacuoles of mouse and human macrophages. Infect. Immun. 68, 3608–3619 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Paetzold, S., Lourido, S., Raupach, B. & Zychlinsky, A. Shigella flexneri phagosomal escape is independent of invasion. Infect. Immun. 75, 4826–4830 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Pilatz, S. et al. Identification of Burkholderia pseudomallei genes required for the intracellular life cycle and in vivo virulence. Infect. Immun. 74, 3576–3586 (2006). This study identifies several B. pseudomallei genes required for vacuole escape and for replication within the cytosol.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Whitworth, T., Popov, V. L., Yu, X. J., Walker, D. H. & Bouyer, D. H. Expression of the Rickettsia prowazekii pld or tlyC gene in Salmonella enterica serovar typhimurium mediates phagosomal escape. Infect. Immun. 73, 6668–6673 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Renesto, P. et al. Identification and characterization of a phospholipase D-superfamily gene in rickettsiae. J. Infect. Dis. 188, 1276–1283 (2003).

    Article  CAS  PubMed  Google Scholar 

  38. Silverman, D. J., Santucci, L. A., Meyers, N. & Sekeyova, Z. Penetration of host cells by Rickettsia rickettsii appears to be mediated by a phospholipase of rickettsial origin. Infect. Immun. 60, 2733–2740 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Winkler, H. H. & Miller, E. T. Phospholipase A and the interaction of Rickettsia prowazekii and mouse fibroblasts (L-929 cells). Infect. Immun. 38, 109–113 (1982).

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Checroun, C., Wehrly, T. D., Fischer, E. R., Hayes, S. F. & Celli, J. Autophagy-mediated reentry of Francisella tularensis into the endocytic compartment after cytoplasmic replication. Proc. Natl Acad. Sci. USA 103, 14578–14583 (2006). This paper reports that F. tularensis subverts the autophagic response by entering and surviving within an FCV following cytosolic replication.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Clemens, D. L., Lee, B. Y. & Horwitz, M. A. Virulent and avirulent strains of Francisella tularensis prevent acidification and maturation of their phagosomes and escape into the cytoplasm in human macrophages. Infect. Immun. 72, 3204–3217 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Santic, M., Molmeret, M., Klose, K. E., Jones, S. & Kwaik, Y. A. The Francisella tularensis pathogenicity island protein IglC and its regulator MglA are essential for modulating phagosome biogenesis and subsequent bacterial escape into the cytoplasm. Cell. Microbiol. 7, 969–979 (2005).

    Article  CAS  PubMed  Google Scholar 

  43. Qin, A., Scott, D. W., Thompson, J. A. & Mann, B. J. Identification of an essential Francisella tularensis subsp. tularensis virulence factor. Infect. Immun. 77, 152–161 (2009).

    Article  CAS  PubMed  Google Scholar 

  44. Yu, J. Inactivation of dsbA, but not dsbC and dsbD, affects the intracellular survival and virulence of Shigella flexneri. Infect. Immun. 66, 3909–3917 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Yu, J., Edwards-Jones, B., Neyrolles, O. & Kroll, J. S. Key role for DsbA in cell-to-cell spread of Shigella flexneri, permitting secretion of Ipa proteins into interepithelial protrusions. Infect. Immun. 68, 6449–6456 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Watarai, M., Tobe, T., Yoshikawa, M. & Sasakawa, C. Disulfide oxidoreductase activity of Shigella flexneri is required for release of Ipa proteins and invasion of epithelial cells. Proc. Natl Acad. Sci. USA 92, 4927–4931 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Stevens, J. M., Galyov, E. E. & Stevens, M. P. Actin-dependent movement of bacterial pathogens. Nature Rev. Microbiol. 4, 91–101 (2006). Useful review that focused on actin-based motility of bacterial pathogens.

    Article  CAS  Google Scholar 

  48. Alberti-Segui, C., Goeden, K. R. & Higgins, D. E. Differential function of Listeria monocytogenes listeriolysin O and phospholipases C in vacuolar dissolution following cell-to-cell spread. Cell. Microbiol. 9, 179–195 (2007).

    Article  CAS  PubMed  Google Scholar 

  49. Yeung, P. S., Na, Y., Kreuder, A. J. & Marquis, H. Compartmentalization of the broad-range phospholipase C activity to the spreading vacuole is critical for Listeria monocytogenes virulence. Infect. Immun. 75, 44–51 (2007).

    Article  CAS  PubMed  Google Scholar 

  50. Schuch, R., Sandlin, R. C. & Maurelli, A. T. A system for identifying post-invasion functions of invasion genes: requirements for the Mxi–Spa type III secretion pathway of Shigella flexneri in intercellular dissemination. Mol. Microbiol. 34, 675–689 (1999).

    Article  CAS  PubMed  Google Scholar 

  51. Page, A. L., Ohayon, H., Sansonetti, P. J. & Parsot, C. The secreted IpaB and IpaC invasins and their cytoplasmic chaperone IpgC are required for intercellular dissemination of Shigella flexneri. Cell. Microbiol. 1, 183–193 (1999).

    Article  CAS  PubMed  Google Scholar 

  52. Monack, D. M. & Theriot, J. A. Actin-based motility is sufficient for bacterial membrane protrusion formation and host cell uptake. Cell. Microbiol. 3, 633–647 (2001).

    Article  CAS  PubMed  Google Scholar 

  53. Goetz, M. et al. Microinjection and growth of bacteria in the cytosol of mammalian host cells. Proc. Natl Acad. Sci. USA 98, 12221–12226 (2001). Provides evidence that the mammalian cell cytosol is restrictive for bacterial growth.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Goebel, W. & Kuhn, M. Bacterial replication in the host cell cytosol. Curr. Opin. Microbiol. 3, 49–53 (2000).

    Article  CAS  PubMed  Google Scholar 

  55. O'Riordan, M. & Portnoy, D. A. The host cytosol: front-line or home front? Trends Microbiol. 10, 361–364 (2002).

    Article  CAS  PubMed  Google Scholar 

  56. Beuzon, C. R. et al. Salmonella maintains the integrity of its intracellular vacuole through the action of SifA. EMBO J. 19, 3235–3249 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Brumell, J. H., Tang, P., Zaharik, M. L. & Finlay, B. B. Disruption of the Salmonella-containing vacuole leads to increased replication of Salmonella enterica serovar Typhimurium in the cytosol of epithelial cells. Infect. Immun. 70, 3264–3270 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Beuzon, C. R., Salcedo, S. P. & Holden, D. W. Growth and killing of a Salmonella enterica serovar Typhimurium sifA mutant strain in the cytosol of different host cell lines. Microbiology 148, 2705–2715 (2002).

    Article  CAS  PubMed  Google Scholar 

  59. Brumell, J. H., Rosenberger, C. M., Gotto, G. T., Marcus, S. L. & Finlay, B. B. SifA permits survival and replication of Salmonella typhimurium in murine macrophages. Cell. Microbiol. 3, 75–84 (2001).

    Article  CAS  PubMed  Google Scholar 

  60. Oyston, P. C. F. Francisella tularensis: unravelling the secrets of an intracellular pathogen. J. Med. Microbiol. 57, 921–930 (2008).

    Article  PubMed  Google Scholar 

  61. Santic, M., Molmeret, M., Klose, K. E. & Abu Kwaik, Y. Francisella tularensis travels a novel, twisted road within macrophages. Trends Microbiol. 14, 37–44 (2006).

    Article  CAS  PubMed  Google Scholar 

  62. Walker, D. H. Rickettsiae and rickettsial infections: the current state of knowledge. Clin. Infect. Dis. 45, S39–S44 (2007).

    Article  Google Scholar 

  63. Cossart, P. & Toledo-Arana, A. Listeria monocytogenes, a unique model in infection biology: an overview. Microbes Infect. 10, 1041–1050 (2008).

    Article  CAS  PubMed  Google Scholar 

  64. Wiersinga, W. J., van der Poll, T., White, N. J., Day, N. P. & Peacock, S. J. Melioidosis: insights into the pathogenicity of Burkholderia pseudomallei. Nature Rev. Microbiol. 4, 272–282 (2006).

    Article  CAS  Google Scholar 

  65. Alberts, B. et al. Molecular Biology of the Cell (Garland Publishing, New York, 2002).

    Google Scholar 

  66. Hwang, C., Sinskey, A. J. & Lodish, H. F. Oxidized redox state of glutathione in the endoplasmic-reticulum. Science 257, 1496–1502 (1992).

    Article  CAS  PubMed  Google Scholar 

  67. Shi, H., Bencze, K. Z., Stemmler, T. L. & Philpott, C. C. A cytosolic iron chaperone that delivers iron to ferritin. Science 320, 1207–1210 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Marquis, H., Bouwer, H. G. A., Hinrichs, D. J. & Portnoy, D. A. Intracytoplasmic growth and virulence of Listeria monocytogenes auxotrophic mutants. Infect. Immun. 61, 3756–3760 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  69. Stritzker, J. et al. Growth, virulence, and immunogenicity of Listeria monocytogenes aro mutants. Infect. Immun. 72, 5622–5629 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Chico-Calero, I. et al. Hpt, a bacterial homolog of the microsomal glucose-6-phosphate translocase, mediates rapid intracellular proliferation in Listeria. Proc. Natl Acad. Sci. USA 99, 431–436 (2002). Together with Reference 83, this study showed that cytosolic bacteria could use hexose phosphates within the cytosol as carbon energy sources.

    Article  CAS  PubMed  Google Scholar 

  71. Eylert, E. et al. Carbon metabolism of Listeria monocytogenes growing inside macrophages. Mol. Microbiol. 69, 1008–1017 (2008).

    Article  CAS  PubMed  Google Scholar 

  72. Ripio, M. T., Brehm, K., Lara, M., Suarez, M. & VazquezBoland, J. A. Glucose-1-phosphate utilization by Listeria monocytogenes is PrfA dependent and coordinately expressed with virulence factors. J. Bacteriol. 179, 7174–7180 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Glaser, P. et al. Comparative genomics of Listeria species. Science 294, 849–852 (2001).

    CAS  PubMed  Google Scholar 

  74. Jordan, S. W. & Cronan, J. E. A new metabolic link. The acyl carrier protein of lipid synthesis donates lipoic acid to the pyruvate dehydrogenase complex in Escherichia coli and mitochondria. J. Biol. Chem. 272, 17903–17906 (1997).

    Article  CAS  PubMed  Google Scholar 

  75. Perham, R. N. Swinging arms and swinging domains in multifunctional enzymes: catalytic machines for multistep reactions. Annu. Rev. Biochem. 69, 961–1004 (2000).

    Article  CAS  PubMed  Google Scholar 

  76. Keeney, K. M., Stuckey, J. A. & O'Riordan, M. X. D. LpIA1-dependent utilization of host lipoyl peptides enables Listeria cytosolic growth and virulence. Mol. Microbiol. 66, 758–770 (2007). Together with Reference 77, this study suggests that L. monocytogenes has evolved to scavenge lipoyl peptides from the mammalian cell cytosol as part of its adaptation to a cytosolic lifestyle.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. O'Riordan, M., Moors, M. A. & Portnoy, D. A. Listeria intracellular growth and virulence require host-derived lipoic acid. Science 302, 462–464 (2003).

    Article  CAS  PubMed  Google Scholar 

  78. Noriega, F. R. et al. Engineered ΔguaB-A ΔvirG Shigella flexneri 2a strain CVD 1205: construction, safety, immunogenicity, and potential efficacy as a mucosal vaccine. Infect. Immun. 64, 3055–3061 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  79. Cersini, A., Martino, M. C., Martini, I., Rossi, G. & Bernardini, M. L. Analysis of virulence and inflammatory potential of Shigella flexneri purine biosynthesis mutants. Infect. Immun. 71, 7002–7013 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Cersini, A., Salvia, A. M. & Bernardini, M. L. Intracellular multiplication and virulence of Shigella flexneri auxotrophic mutants. Infect. Immun. 66, 549–557 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  81. Bartoleschi, C. et al. Selection of Shigella flexneri candidate virulence genes specifically induced in bacteria resident in host cell cytoplasm. Cell. Microbiol. 4, 613–626 (2002).

    Article  CAS  PubMed  Google Scholar 

  82. Lucchini, S., Liu, H., Jin, Q., Hinton, J. C. D. & Yu, J. Transcriptional adaptation of Shigella flexneri during infection of macrophages and epithelial cells: insights into the strategies of a cytosolic bacterial pathogen. Infect. Immun. 73, 88–102 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Runyen-Janecky, L. J. & Payne, S. M. Identification of chromosomal Shigella flexneri genes induced by the eukaryotic intracellular environment. Infect. Immun. 70, 4379–4388 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Austin, F. E. & Winkler, H. H. Proline incorporation into protein by Rickettsia prowazekii during growth in chinese hamster ovary (Cho-K1) cells. Infect. Immun. 56, 3167–3172 (1988).

    CAS  PubMed  PubMed Central  Google Scholar 

  85. Austin, F. E., Turco, J. & Winkler, H. H. Rickettsia prowazekii requires host cell serine and glycine for growth. Infect. Immun. 55, 240–244 (1987).

    CAS  PubMed  PubMed Central  Google Scholar 

  86. Renesto, P., Ogata, H., Audic, S., Claverie, J. M. & Raoult, D. Some lessons from Rickettsia genomics. FEMS Microbiol. Rev. 29, 99–117 (2005).

    Article  CAS  PubMed  Google Scholar 

  87. Fuller, J. R. et al. RipA, a cytoplasmic membrane protein conserved among Francisella species, is required for intracellular survival. Infect. Immun. 76, 4934–4943 (2008). Description of a protein conserved amongst the Francisella species that is required for intracellular replication within the cell cytosol.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Santic, M. et al. A Francisella tularensis pathogenicity island protein essential for bacterial proliferation within the host cell cytosol. Cell. Microbiol. 9, 2391–2403 (2007).

    Article  CAS  PubMed  Google Scholar 

  89. Bonquist, L., Lindgren, H., Golovliov, I., Guina, T. & Sjostedt, A. MglA and Igl proteins contribute to the modulation of Francisella tularensis live vaccine strain-containing phagosomes in murine macrophages. Infect. Immun. 76, 3502–3510 (2008).

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  90. Hiemstra, P. S., van den Barselaar, M. T., Roest, M., Nibbering, P. H. & van Furth, R. Ubiquicidin, a novel murine microbicidal protein present in the cytosolic fraction of macrophages. J. Leukoc. Biol. 66, 423–428 (1999).

    Article  CAS  PubMed  Google Scholar 

  91. Delbridge, L. M. & O'Riordan, M. X. Innate recognition of intracellular bacteria. Curr. Opin. Immunol. 19, 10–16 (2007).

    Article  CAS  PubMed  Google Scholar 

  92. Kufer, T. A. & Sansonetti, P. J. Sensing of bacteria: NOD a lonely job. Curr. Opin. Microbiol. 10, 62–69 (2007).

    Article  CAS  PubMed  Google Scholar 

  93. Orvedahl, A. & Levine, B. Eating the enemy within: autophagy in infectious diseases. Cell Death Differ. 16, 57–69 (2008).

    Article  PubMed  CAS  Google Scholar 

  94. Xie, Z. & Klionsky, D. J. Autophagosome formation: core machinery and adaptations. Nature Cell Biol. 9, 1102–1109 (2007).

    Article  CAS  PubMed  Google Scholar 

  95. Zhao, Z. et al. Autophagosome-independent essential function for the autophagy protein Atg5 in cellular immunity to intracellular pathogens. Cell Host Microbe 4, 458–469 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  96. Rich, K. A., Burkett, C. & Webster, P. Cytoplasmic bacteria can be targets for autophagy. Cell. Microbiol. 5, 455–468 (2003).

    Article  CAS  PubMed  Google Scholar 

  97. Yano, T. et al. Autophagic control of listeria through intracellular innate immune recognition in drosophila. Nature Immunol. 9, 908–916 (2008).

    Article  CAS  Google Scholar 

  98. Py, B. F., Lipinski, M. M. & Yuan, J. Y. Autophagy limits Listeria monocytogenes intracellular growth in the early phase of primary infection. Autophagy 3, 117–125 (2007).

    Article  CAS  PubMed  Google Scholar 

  99. Birmingham, C. L. et al. Listeria monocytogenes evades killing by autophagy during colonization of host cells. Autophagy 3, 442–451 (2007).

    Article  CAS  PubMed  Google Scholar 

  100. Birmingham, C. L. et al. Listeriolysin O allows Listeria monocytogenes replication in macrophage vacuoles. Nature 451, 350–354 (2008). This paper suggests that LLO enables L. monocytogenes to establish a persistent infection of macrophages by allowing survival within a specialized vacuole.

    Article  CAS  PubMed  Google Scholar 

  101. Ogawa, M. et al. Escape of intracellular Shigella from autophagy. Science 307, 727–731 (2005). This paper describes how S. flexneri evades autophagy by secreting the type III secretion system effector IcsB.

    Article  CAS  PubMed  Google Scholar 

  102. Cullinane, M. et al. Stimulation of autophagy suppresses the intracellular survival of Burkholderia pseudomallei in mammalian cell lines. Autophagy 4, 744–753 (2008).

    Article  CAS  PubMed  Google Scholar 

  103. Stevens, M. P. et al. Attenuated virulence and protective efficacy of a Burkholderia pseudomallei bsa type III secretion mutant in murine models of melioidosis. Microbiology 150, 2669–2676 (2004).

    Article  CAS  PubMed  Google Scholar 

  104. Butchar, J. P. et al. Microarray analysis of human monocytes infected with Francisella tularensis identifies new targets of host response subversion. PLoS ONE 3, e2924 (2008).

    Article  PubMed  PubMed Central  Google Scholar 

  105. Stamm, L. M. et al. Mycobacterium marinum escapes from phagosomes and is propelled by actin-based motility. J. Exp. Med. 198, 1361–1368 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  106. van der Wel, N. et al. M. tuberculosis and M. leprae translocate from the phagolysosome to the cytosol in myeloid cells. Cell 129, 1287–1298 (2007).

    Article  CAS  PubMed  Google Scholar 

  107. Tala, A. et al. The HrpB–HrpA two-partner secretion system is essential for intracellular survival of Neisseria meningitidis. Cell. Microbiol. 10, 2461–2482 (2008).

    Article  CAS  PubMed  Google Scholar 

  108. Gouin, E., Welch, M. D. & Cossart, P. Actin-based motility of intracellular pathogens. Curr. Opin. Microbiol. 8, 35–45 (2005).

    Article  CAS  PubMed  Google Scholar 

  109. Paavilainen, V. O., Bertling, E., Falck, S. & Lappalainen, P. Regulation of cytoskeletal dynamics by actin-monomer-binding proteins. Trends Cell Biol. 14, 386–394 (2004).

    Article  CAS  PubMed  Google Scholar 

  110. Welch, M. D. & Mullins, R. D. Cellular control of actin nucleation. Annu. Rev. Cell Dev. Biol. 18, 247–288 (2002).

    Article  CAS  PubMed  Google Scholar 

  111. Kocks, C. et al. L. monocytogenes-induced actin assembly requires the actA gene product, a surface protein. Cell 68, 521–531 (1992).

    Article  CAS  PubMed  Google Scholar 

  112. Stevens, M. P. et al. Identification of a bacterial factor required for actin-based motility of Burkholderia pseudomallei. Mol. Microbiol. 56, 40–53 (2005).

    Article  CAS  PubMed  Google Scholar 

  113. Gouin, E. et al. The RickA protein of Rickettsia conorii activates the Arp2/3 complex. Nature 427, 457–461 (2004).

    Article  CAS  PubMed  Google Scholar 

  114. Jeng, R. L. et al. A Rickettsia WASP-like protein activates the Arp2/3 complex and mediates actin-based motility. Cell. Microbiol. 6, 761–769 (2004).

    Article  CAS  PubMed  Google Scholar 

  115. Boujemaa-Paterski, R. et al. Listeria protein ActA mimics WASP family proteins: it activates filament barbed end branching by Arp2/3 complex. Biochemistry 40, 11390–11404 (2001).

    Article  CAS  PubMed  Google Scholar 

  116. Welch, M. D., Rosenblatt, J., Skoble, J., Portnoy, D. A. & Mitchison, T. J. Interaction of human Arp2/3 complex and the Listeria monocytogenes ActA protein in actin filament nucleation. Science 281, 105–108 (1998).

    Article  CAS  PubMed  Google Scholar 

  117. Breitbach, K. et al. Actin-based motility of Burkholderia pseudomallei involves the Arp 2/3 complex, but not N-WASP and Ena/VASP proteins. Cell. Microbiol. 5, 385–393 (2003).

    Article  CAS  PubMed  Google Scholar 

  118. Bernardini, M. L., Mounier, J., Dhauteville, H., Coquisrondon, M. & Sansonetti, P. J. Identification of icsA, a plasmid locus of Shigella flexneri that governs bacterial intra- and intercellular spread through interaction with F-actin. Proc. Natl Acad. Sci. USA 86, 3867–3871 (1989).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  119. Egile, C. et al. Activation of the CDC42 effector N-WASP by the Shigella flexneri IcsA protein promotes actin nucleation by Arp2/3 complex and bacterial actin-based motility. J. Cell Biol. 146, 1319–1332 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  120. Van Kirk, L. S., Hayes, S. F. & Heinzen, R. A. Ultrastructure of Rickettsia rickettsii actin tails and localization of cytoskeletal proteins. Infect. Immun. 68, 4706–4713 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  121. Stevens, M. P. et al. An Inv/Mxi-Spa-like type III protein secretion system in Burkholderia pseudomallei modulates intracellular behaviour of the pathogen. Mol. Microbiol. 46, 649–659 (2002).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the European Initiative for Basic Research in Microbiology and Infectious Diseases grant D005-P09205 funded by the Commission of the European Communities (to K.R.) and by the Fondation pour la Recherche Médicale (to B.M.).

Author information

Authors and Affiliations

  1. Department of Microbiology, Centre for Molecular Microbiology and Infection, Flowers Building, Armstrong Road, Imperial College London, London, SW7 2AZ, UK

    Katrina Ray, Benoit Marteyn & Christoph M. Tang

  2. Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, 28 rue du Dr Roux, F -75724, Paris, Cédex 15, France

    Benoit Marteyn & Philippe J. Sansonetti

Authors
  1. Katrina Ray
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Benoit Marteyn
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Philippe J. Sansonetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christoph M. Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Christoph M. Tang.

Related links

Related links

DATABASES

Entrez Genome Project

Bacillus subtilis

Burkholderia pseudomallei

Drosophila melanogaster

Escherichia coli

Francisella tularensis

Listeria monocytogenes

Mycobacterium marinum

Neisseria meningitidis

Rickettsia conorii

Rickettsia prowazekii

Salmonella enterica subspecies enterica serovar Typhimurium

Shigella flexneri

Yersinia enterocolitica

Yersinia pseudotuberculosis

FURTHER INFORMATION

Christoph M. Tang's homepage

Glossary

Trigger mechanism

A mechanism used by bacteria, such as the genera Shigella and Salmonella, to enter cells. Bacteria interact directly with the eukaryotic cell cytoskeleton by injecting bacterial effectors through a dedicated secretion system. These effectors cause massive cytoskeletal rearrangements to engulf the bacterium in an entry vacuole.

Zipper mechanism

A mechanism used by bacteria, such as the genera Yersinia or Listeria, to enter cells. Bacteria contact and adhere to the eukaryotic cell through the binding of a bacterial surface protein to a eukaryotic surface receptor, often a transmembrane cell-adhesion protein. Modest membrane extensions and cytoskeletal rearrangements engulf the bacterium in an entry vacuole.

Vacuole

A single-membrane organelle within the cell cytosol that encloses a fluid-filled compartment.

Phagolysosome

A membrane-enclosed organelle formed by the fusion of a lysosome, which is an organelle containing hydrolytic enzymes, and a phagosome, which is a membranous vacuole formed around a particle.

Lysosome

A membrane-bound organelle that contains hydrolytic enzymes.

Microbicidal

An activity that is lethal for microorganisms.

Listeriolysin O

(LLO). A thiol-activated cholesterol-dependent pore-forming toxin produced by Listeria monocytogenes. The production of LLO is essential for escape of the bacterium from the vacuole into the cytosol during invasion, and therefore LLO is a key virulence factor.

Type C phospholipase

A subclass of enzymes that cleave the polar head group of phosphoinositides between the glycerol and phosphate moieties.

Thiol reductase

An enzyme that catalyses disulphide bond reduction.

Type III secretion system

A secretion apparatus of Gram-negative bacteria that allows bacterial effector proteins to be delivered directly into the eukaryotic cell cytosol through a bacterial molecular needle complex.

Oxidoreductase

A class of enzymes that catalyse oxidoreduction reactions which transfer electrons from a hydrogen donor to a hydrogen acceptor.

Disulphide bond

A single covalent bond formed from the coupling of two thiol groups, the functional group of which is composed of a sulphur and a hydrogen atom.

Glutathione

A tripeptide that acts as an antioxidant and electron donor in the cell cytosol by reducing disulphide bonds formed between cysteines of cytoplasmic proteins.

Ferritin

A globular protein that consists of 24 subunits and is the main intracellular iron storage protein in eukaryotes.

Auxotrophic

The state of an organism when it is unable to synthesize a particular organic compound required for growth.

Diaminopimelate

The ionic form of the amino acid diaminopimelic acid, which is a compound found in the peptidoglycan of bacteria.

Oxygen tension

The partial pressure of oxygen.

Complement

A crucial part of the innate immune system that consists of enzyme cascades which lead to bacterial lysis and promote phagocytosis.

Antimicrobial peptide

A conserved component of the innate immune system that consists of polypeptides, with fewer than 100 amino acid residues, that have the ability to kill microorganisms.

Pathogen-associated molecular pattern

A small molecular motif that is consistently found on pathogens and is recognized as a non-self molecule by a pattern recognition receptor of the innate immune system.

Nod-like receptor

One of a family of proteins that serve as pattern recognition receptors, in that they sense microbial motifs in the cell cytoplasm.

Autophagy

A degradative pathway by which cytosolic content, organelles and pathogens are delivered to lysosomes as part of cellular homeostasis and innate immunity.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ray, K., Marteyn, B., Sansonetti, P. et al. Life on the inside: the intracellular lifestyle of cytosolic bacteria. Nat Rev Microbiol 7, 333–340 (2009). https://doi.org/10.1038/nrmicro2112

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nrmicro2112

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing