Home LiteratureArticle Details
PMID: 10768992 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Intracellular growth of Legionella pneumophila in Dictyostelium discoideum, a system for genetic analysis of host-pathogen interactions.

Infection and immunity ·Vol. 68 ·No. 5 ·2000-05-00 ·Pages 2939-47

Solomon JM, Rupper A, Cardelli JA, Isberg RR

Abstract

Conditions were established in which Legionella pneumophila, an intracellular bacterial pathogen, could replicate within the unicellular organism Dictyostelium discoideum. By several criteria, L. pneumophila grew by the same mechanism within D. discoideum as it does in amoebae and macrophages. Bacteria grew within membrane-bound vesicles associated with rough endoplasmic reticulum, and L. pneumophila dot/icm mutants, blocked for growth in macrophages and amoebae, also did not grow in D. discoideum. Internalized L. pneumophila avoided degradation by D. discoideum and showed evidence of reduced fusion with endocytic compartments. The ability of L. pneumophila to grow within D. discoideum depended on the growth state of the cells. D. discoideum grown as adherent monolayers was susceptible to L. pneumophila infection and to contact-dependent cytotoxicity during high-multiplicity infections, whereas D. discoideum grown in suspension was relatively resistant to cytotoxicity and did not support intracellular growth. Some known D. discoideum mutants were examined for their effect on growth of L. pneumophila. The coronin mutant and the myoA/B double myosin I mutant were more permissive than wild-type strains for intracellular growth. Growth of L. pneumophila in a G(beta) mutant was slightly reduced compared to the parent strain. This work demonstrates the usefulness of the L. pneumophila-D. discoideum system for genetic analysis of host-pathogen interactions.

MeSH Terms
Animals Culture Media Dictyostelium/genetics,microbiology Genes, Bacterial Intracellular Fluid/microbiology Intracellular Membranes Legionella pneumophila/genetics,growth & development Lysosomes Membrane Proteins
Chemicals
Culture Media Membrane Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Solomon J M
Howard Hughes Medical Institute, Department of Molecular Biology and Microbiology, Tufts University Medical School, Boston, Massachusetts 02111, USA.
Rupper A
Cardelli J A
Isberg R R
References (55)
55 references, click to expand
  1. Two distinct defects in intracellular growth complemented by a single genetic locus in Legionella pneumophila.
    Mol Microbiol. 1993 Jan;7(1):7-19 PMID: 8382332
  2. Dictyostelium mutants lacking the cytoskeletal protein coronin are defective in cytokinesis and cell motility.
    J Cell Biol. 1993 Jan;120(1):163-73 PMID: 8380174
  3. Vacuolar H(+)-ATPase of Dictyostelium discoideum. A monoclonal antibody study.
    J Cell Sci. 1993 Dec;106 ( Pt 4):1103-13 PMID: 8126094
  4. Characterization of lysosomal membrane proteins of Dictyostelium discoideum. A complex population of acidic integral membrane glycoproteins, Rab GTP-binding proteins and vacuolar ATPase subunits.
    J Biol Chem. 1994 Oct 14;269(41):25719-27 PMID: 7929276
  5. The Legionella pneumophila icm locus: a set of genes required for intracellular multiplication in human macrophages.
    Mol Microbiol. 1994 Nov;14(4):797-808 PMID: 7891565
  6. Altered intracellular targeting properties associated with mutations in the Legionella pneumophila dotA gene.
    Mol Microbiol. 1994 Nov;14(4):809-22 PMID: 7891566
  7. The G protein beta subunit is essential for multiple responses to chemoattractants in Dictyostelium.
    J Cell Biol. 1995 Jun;129(6):1667-75 PMID: 7790362
  8. Association of Legionella pneumophila with the macrophage endoplasmic reticulum.
    Infect Immun. 1995 Sep;63(9):3609-20 PMID: 7642298
  9. A phosphatidylinositol (PI) kinase gene family in Dictyostelium discoideum: biological roles of putative mammalian p110 and yeast Vps34p PI 3-kinase homologs during growth and development.
    Mol Cell Biol. 1995 Oct;15(10):5645-56 PMID: 7565716
  10. Integrated maps of the chromosomes in Dictyostelium discoideum.
    Genetics. 1995 Sep;141(1):147-57 PMID: 8536963
  11. Dictyostelium myosin I double mutants exhibit conditional defects in pinocytosis.
    J Cell Biol. 1995 Dec;131(5):1205-21 PMID: 8522584
  12. Coronin involved in phagocytosis: dynamics of particle-induced relocalization visualized by a green fluorescent protein Tag.
    Cell. 1995 Dec 15;83(6):915-24 PMID: 8521515
  13. Identification of Legionella pneumophila mutants that have aberrant intracellular fates.
    Infect Immun. 1996 Jul;64(7):2585-94 PMID: 8698483
  14. FACS-optimized mutants of the green fluorescent protein (GFP).
    Gene. 1996;173(1 Spec No):33-8 PMID: 8707053
  15. The dorsoventral regulatory gene cassette spätzle/Toll/cactus controls the potent antifungal response in Drosophila adults.
    Cell. 1996 Sep 20;86(6):973-83 PMID: 8808632
  16. The molecular ecology of legionellae.
    Trends Microbiol. 1996 Jul;4(7):286-90 PMID: 8829338
  17. Evidence for apoptosis of human macrophage-like HL-60 cells by Legionella pneumophila infection.
    Infect Immun. 1996 Dec;64(12):4900-6 PMID: 8945524
  18. Use of salt to isolate Legionella pneumophila mutants unable to replicate in macrophages.
    Ann N Y Acad Sci. 1996 Oct 25;797:271-2 PMID: 8993377
  19. Fluid-phase uptake by macropinocytosis in Dictyostelium.
    J Cell Sci. 1997 Jan;110 ( Pt 2):105-12 PMID: 9044041
  20. Phagosomal proteins of Dictyostelium discoideum.
    J Eukaryot Microbiol. 1997 Jul-Aug;44(4):284-92 PMID: 9225442
  21. Evidence for a recycling role for Rab7 in regulating a late step in endocytosis and in retention of lysosomal enzymes in Dictyostelium discoideum.
    Mol Biol Cell. 1997 Jul;8(7):1343-60 PMID: 9243512
  22. Utilization of similar mechanisms by Legionella pneumophila to parasitize two evolutionarily distant host cells, mammalian macrophages and protozoa.
    Infect Immun. 1997 Nov;65(11):4738-46 PMID: 9353059
  23. The Legionella pneumophila icmGCDJBF genes are required for killing of human macrophages.
    Infect Immun. 1998 May;66(5):2245-55 PMID: 9573114
  24. Legionella pneumophila DotA protein is required for early phagosome trafficking decisions that occur within minutes of bacterial uptake.
    Mol Microbiol. 1998 May;28(3):663-74 PMID: 9632267
  25. Expression of Legionella pneumophila virulence traits in response to growth conditions.
    Infect Immun. 1998 Jul;66(7):3029-34 PMID: 9632562
  26. G protein beta subunit-null mutants are impaired in phagocytosis and chemotaxis due to inappropriate regulation of the actin cytoskeleton.
    J Cell Biol. 1998 Jun 29;141(7):1529-37 PMID: 9647646
  27. Early events in phagosome establishment are required for intracellular survival of Legionella pneumophila.
    Infect Immun. 1998 Sep;66(9):4450-60 PMID: 9712800
  28. Role of PKA in the timing of developmental events in Dictyostelium cells.
    Microbiol Mol Biol Rev. 1998 Sep;62(3):684-94 PMID: 9729606
  29. Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis.
    Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):715-20 PMID: 9892699
  30. The Toll-receptor family and control of innate immunity.
    Curr Opin Immunol. 1999 Feb;11(1):13-8 PMID: 10047546
  31. Taking the plunge. Terminal differentiation in Dictyostelium.
    Trends Genet. 1999 Jan;15(1):15-9 PMID: 10087928
  32. Legionella pneumophila utilizes the same genes to multiply within Acanthamoeba castellanii and human macrophages.
    Infect Immun. 1999 May;67(5):2117-24 PMID: 10225863
  33. A coat protein on phagosomes involved in the intracellular survival of mycobacteria.
    Cell. 1999 May 14;97(4):435-47 PMID: 10338208
  34. Characterization of a new region required for macrophage killing by Legionella pneumophila.
    Infect Immun. 1997 Dec;65(12):5057-66 PMID: 9393796
  35. Conjugative transfer by the virulence system of Legionella pneumophila.
    Science. 1998 Feb 6;279(5352):873-6 PMID: 9452389
  36. Host cell killing and bacterial conjugation require overlapping sets of genes within a 22-kb region of the Legionella pneumophila genome.
    Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1669-74 PMID: 9465074
  37. Evidence for pore-forming ability by Legionella pneumophila.
    Mol Microbiol. 1998 Jan;27(2):323-36 PMID: 9484888
  38. Identification of linked Legionella pneumophila genes essential for intracellular growth and evasion of the endocytic pathway.
    Infect Immun. 1998 Mar;66(3):950-8 PMID: 9488381
  39. Sensitivity of Dictyostelium discoideum to nucleic acid analogues.
    Exp Cell Res. 1971 Feb;64(2):484-6 PMID: 5542656
  40. Genetics of growth in axenic medium of the cellular slime mould Dictyostelium discoideum.
    Nature. 1974 Jan 18;247(5437):142-3 PMID: 4855676
  41. Demonstration of the agent of Legionnaires' disease in tissue.
    N Engl J Med. 1977 Dec 1;297(22):1218-20 PMID: 72358
  42. Charcoal-yeast extract agar: primary isolation medium for Legionella pneumophila.
    J Clin Microbiol. 1979 Oct;10(4):437-41 PMID: 393713
  43. Legionnaires' disease bacterium (Legionella pneumophila) multiples intracellularly in human monocytes.
    J Clin Invest. 1980 Sep;66(3):441-50 PMID: 7190579
  44. Formation of a novel phagosome by the Legionnaires' disease bacterium (Legionella pneumophila) in human monocytes.
    J Exp Med. 1983 Oct 1;158(4):1319-31 PMID: 6619736
  45. The Legionnaires' disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes.
    J Exp Med. 1983 Dec 1;158(6):2108-26 PMID: 6644240
  46. Legionella pneumophila inhibits acidification of its phagosome in human monocytes.
    J Cell Biol. 1984 Dec;99(6):1936-43 PMID: 6501409
  47. Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination.
    Science. 1987 May 29;236(4805):1086-91 PMID: 3576222
  48. Endocytosis and recognition mechanisms in Dictyostelium discoideum.
    Methods Cell Biol. 1987;28:129-37 PMID: 3600403
  49. Cultivation and synchronous morphogenesis of Dictyostelium under controlled experimental conditions.
    Methods Cell Biol. 1987;28:9-29 PMID: 3298997
  50. IFN-gamma-activated human alveolar macrophages inhibit the intracellular multiplication of Legionella pneumophila.
    J Immunol. 1988 Jun 1;140(11):3978-81 PMID: 3131422
  51. Electrophoretic karyotype for Dictyostelium discoideum.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8247-51 PMID: 2236037
  52. A series of wide-host-range low-copy-number vectors that allow direct screening for recombinants.
    Gene. 1991 Jan 2;97(1):39-47 PMID: 1847347
  53. Techniques for the visualisation of cytoskeletal components in Dictyostelium discoideum.
    Electron Microsc Rev. 1991;4(2):343-76 PMID: 1932587
  54. Myosin IB null mutants of Dictyostelium exhibit abnormalities in motility.
    Cell Motil Cytoskeleton. 1991;20(4):301-15 PMID: 1666340
  55. The unconventional myosin encoded by the myoA gene plays a role in Dictyostelium motility.
    Mol Biol Cell. 1993 Feb;4(2):233-46 PMID: 8382977
Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.68.5.2939-2947.2000
Published
2000-05-00
Pages
2939-47
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC97507
Subset
IM
Grants
NIDDK NIH HHS · DK39232 · United States
External Links
PubMed source DOI: 10.1128/IAI.68.5.2939-2947.2000 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2