Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Feb;57(2):477–486. doi: 10.1128/iai.57.2.477-486.1989

Intracellular spread of Shigella flexneri associated with the kcpA locus and a 140-kilodalton protein.

T Pál 1, J W Newland 1, B D Tall 1, S B Formal 1, T L Hale 1
PMCID: PMC313121  PMID: 2643571

Abstract

Escherichia coli K-12 hybrids carrying both the 220-kilobase plasmid and the purE-linked kcpA locus from Shigella flexneri expressed a 140-kilodalton (kDa) protein which was recognized by convalescent sera from monkeys infected with S. flexneri. These hybrids were tested for the ability to produce plaques in HeLa cell monolayers. Hybrid strains which carried both the 220-kilobase plasmid and the kcpA locus had a plaque-forming efficiency of at least 10(-4) PFU/CFU, whereas the plaque-forming efficiency of hybrids that carried only the shigella invasion plasmid ranged from undetectable to 10(-6). Variants were purified from the rare plaques formed by E. coli hybrids that carried only the shigella invasion plasmid. These plaque-purified strains also expressed the 140-kDa protein, and they had a plaque-forming efficiency of at least 10(-4). Transduction of the purE locus from a plaque-purified hybrid into a non-plaque-forming E. coli K-12 strain did not alter the phenotype of the recipient, but conjugation of the shigella invasion plasmid into this transductant reconstituted both expression of the 140-kDa protein and the plaque-forming phenotype. Invasive E. coli K-12 hybrids carrying only the shigella invasion plasmid remained localized within discrete areas of the HeLa cell cytoplasm, whereas hybrids that also carried the S. flexneri kcpA locus grew in a dispersed pattern throughout the host cell cytoplasm. The dispersal of these organisms was inhibited by cytochalasin D, which suggested that host cell microfilaments may play a role in the intracellular spread of enteroinvasive pathogens.

Full text

PDF
477

Images in this article

479Image
on p.479
480Image
on p.480
480Image
on p.480
480Image
on p.480
481Image
on p.481
482Image
on p.482
483Image
on p.483
484Image
on p.484

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams R. J., Pollard T. D. Propulsion of organelles isolated from Acanthamoeba along actin filaments by myosin-I. Nature. 1986 Aug 21;322(6081):754–756. doi: 10.1038/322754a0. [DOI] [PubMed] [Google Scholar]
  2. Baudry B., Maurelli A. T., Clerc P., Sadoff J. C., Sansonetti P. J. Localization of plasmid loci necessary for the entry of Shigella flexneri into HeLa cells, and characterization of one locus encoding four immunogenic polypeptides. J Gen Microbiol. 1987 Dec;133(12):3403–3413. doi: 10.1099/00221287-133-12-3403. [DOI] [PubMed] [Google Scholar]
  3. Buysse J. M., Stover C. K., Oaks E. V., Venkatesan M., Kopecko D. J. Molecular cloning of invasion plasmid antigen (ipa) genes from Shigella flexneri: analysis of ipa gene products and genetic mapping. J Bacteriol. 1987 Jun;169(6):2561–2569. doi: 10.1128/jb.169.6.2561-2569.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dinari G., Hale T. L., Austin S. W., Formal S. B. Local and systemic antibody responses to Shigella infection in rhesus monkeys. J Infect Dis. 1987 May;155(5):1065–1069. doi: 10.1093/infdis/155.5.1065. [DOI] [PubMed] [Google Scholar]
  5. Formal S. B., Gemski P., Baron L. S., Labrec E. H. Genetic Transfer of Shigella flexneri Antigens to Escherichia coli K-12. Infect Immun. 1970 Mar;1(3):279–287. doi: 10.1128/iai.1.3.279-287.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Formal S. B., Hale T. L., Kapfer C., Cogan J. P., Snoy P. J., Chung R., Wingfield M. E., Elisberg B. L., Baron L. S. Oral vaccination of monkeys with an invasive Escherichia coli K-12 hybrid expressing Shigella flexneri 2a somatic antigen. Infect Immun. 1984 Nov;46(2):465–469. doi: 10.1128/iai.46.2.465-469.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Griffiths E., Stevenson P., Hale T. L., Formal S. B. Synthesis of aerobactin and a 76,000-dalton iron-regulated outer membrane protein by Escherichia coli K-12-Shigella flexneri hybrids and by enteroinvasive strains of Escherichia coli. Infect Immun. 1985 Jul;49(1):67–71. doi: 10.1128/iai.49.1.67-71.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hale T. L., Formal S. B. Protein synthesis in HeLa or Henle 407 cells infected with Shigella dysenteriae 1, Shigella flexneri 2a, or Salmonella typhimurium W118. Infect Immun. 1981 Apr;32(1):137–144. doi: 10.1128/iai.32.1.137-144.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hale T. L. Invasion of epithelial cells by shigellae. Ann Inst Pasteur Microbiol. 1986 May-Jun;137A(3):311–314. doi: 10.1016/s0769-2609(86)80040-0. [DOI] [PubMed] [Google Scholar]
  10. Hale T. L., Morris R. E., Bonventre P. F. Shigella infection of henle intestinal epithelial cells: role of the host cell. Infect Immun. 1979 Jun;24(3):887–894. doi: 10.1128/iai.24.3.887-894.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hale T. L., Oaks E. V., Formal S. B. Identification and antigenic characterization of virulence-associated, plasmid-coded proteins of Shigella spp. and enteroinvasive Escherichia coli. Infect Immun. 1985 Dec;50(3):620–629. doi: 10.1128/iai.50.3.620-629.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hale T. L., Sansonetti P. J., Schad P. A., Austin S., Formal S. B. Characterization of virulence plasmids and plasmid-associated outer membrane proteins in Shigella flexneri, Shigella sonnei, and Escherichia coli. Infect Immun. 1983 Apr;40(1):340–350. doi: 10.1128/iai.40.1.340-350.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Labrec E. H., Schneider H., Magnani T. J., Formal S. B. EPITHELIAL CELL PENETRATION AS AN ESSENTIAL STEP IN THE PATHOGENESIS OF BACILLARY DYSENTERY. J Bacteriol. 1964 Nov;88(5):1503–1518. doi: 10.1128/jb.88.5.1503-1518.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Makino S., Sasakawa C., Kamata K., Kurata T., Yoshikawa M. A genetic determinant required for continuous reinfection of adjacent cells on large plasmid in S. flexneri 2a. Cell. 1986 Aug 15;46(4):551–555. doi: 10.1016/0092-8674(86)90880-9. [DOI] [PubMed] [Google Scholar]
  15. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Matteoni R., Kreis T. E. Translocation and clustering of endosomes and lysosomes depends on microtubules. J Cell Biol. 1987 Sep;105(3):1253–1265. doi: 10.1083/jcb.105.3.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maurelli A. T., Baudry B., d'Hauteville H., Hale T. L., Sansonetti P. J. Cloning of plasmid DNA sequences involved in invasion of HeLa cells by Shigella flexneri. Infect Immun. 1985 Jul;49(1):164–171. doi: 10.1128/iai.49.1.164-171.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miranda A. F., Godman G. C., Deitch A. D., Tanenbaum S. W. Action of cytochalasin D on cells of established lines. I. Early events. J Cell Biol. 1974 May;61(2):481–500. doi: 10.1083/jcb.61.2.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Oaks E. V., Hale T. L., Formal S. B. Serum immune response to Shigella protein antigens in rhesus monkeys and humans infected with Shigella spp. Infect Immun. 1986 Jul;53(1):57–63. doi: 10.1128/iai.53.1.57-63.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oaks E. V., Wingfield M. E., Formal S. B. Plaque formation by virulent Shigella flexneri. Infect Immun. 1985 Apr;48(1):124–129. doi: 10.1128/iai.48.1.124-129.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ogawa H., Nakamura A., Nakaya R. Cinemicrographic study of tissue cell cultures infected with Shigella flexneri. Jpn J Med Sci Biol. 1968 Aug;21(4):259–273. doi: 10.7883/yoken1952.21.259. [DOI] [PubMed] [Google Scholar]
  22. Sakai T., Sasakawa C., Yoshikawa M. Expression of four virulence antigens of Shigella flexneri is positively regulated at the transcriptional level by the 30 kiloDalton virF protein. Mol Microbiol. 1988 Sep;2(5):589–597. doi: 10.1111/j.1365-2958.1988.tb00067.x. [DOI] [PubMed] [Google Scholar]
  23. Sansonetti P. J., Hale T. L., Dammin G. J., Kapfer C., Collins H. H., Jr, Formal S. B. Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri. Infect Immun. 1983 Mar;39(3):1392–1402. doi: 10.1128/iai.39.3.1392-1402.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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. 1986 Feb;51(2):461–469. doi: 10.1128/iai.51.2.461-469.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sasakawa C., Kamata K., Sakai T., Makino S., Yamada M., Okada N., Yoshikawa M. Virulence-associated genetic regions comprising 31 kilobases of the 230-kilobase plasmid in Shigella flexneri 2a. J Bacteriol. 1988 Jun;170(6):2480–2484. doi: 10.1128/jb.170.6.2480-2484.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sasakawa C., Makino S., Kamata K., Yoshikawa M. Isolation, characterization, and mapping of Tn5 insertions into the 140-megadalton invasion plasmid defective in the mouse Sereny test in Shigella flexneri 2a. Infect Immun. 1986 Oct;54(1):32–36. doi: 10.1128/iai.54.1.32-36.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES