Molecular Genetic Analysis of Virulence Factors of Shigella Flexneri

  • Masanosuke Yoshikawa
  • Chihiro Sasakawa
  • Takashi Sakai
  • Masatoshi Yamada
  • Nobuhiko Okada
  • Sou-ichi Makino


Labrec et al.7 established in 1964 that invasion into epithelial cells was the essential early step in the pathogenesis of bacillary dysentery. The virulent translucent strain of Shigella flexneri invaded the intestinal epithelium, while the avirulent opaque variant did not, in spite of the finding, that both had the same LD50 upon intraperitoneal injection. Ogawa et al.15 performed similar experiments and obtained essentially the same results. However, they made one important additional observation; within 24 hours of infection, the bacteria existed exclusively in the epithelial cells without direct penetration into the lamina propria and bacterial multiplication therein. Thus, virulent shigellae invade the colonic epithelium and spread successively to adjacent epithelial cells resulting in a severe inflammatory reaction, further spread of the bacteria to deeper tissue and, ultimately, to ulceration and bloody diarrhea.


Corneal Epithelium Bacillary Dysentery Shigella Flexneri Adjacent Epithelial Cell Intergeneric Conjugation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Buysse JM, Stover CK, Oaks, EV, Venkatesan M, Kopecko DJ, 1987, Molecular cloning of invasion plasmid antigen (ipa) genes from Shigella flexneri: analysis of ipa gene products and genetic mapping, J Bacteriol., 169:2561–2569.Google Scholar
  2. 2.
    Casadaban MJ, Chou J, Cohen SN, 1980, In vitro gene fusions that join an enzymatically active (3-galactosidase segment to amino - terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals, J Bacteriol., 143:971–980.Google Scholar
  3. 3.
    Close TJ, Rodriguez RL, 1982, Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal element, Gene, 20:305–316.CrossRefGoogle Scholar
  4. 4.
    Formal SB, Gemski P Jr, Baron LS, LaBrec EH, 1970, Genetic transfer of Shigella flexneri antigens to Escherichia coli K-12, Infect Immun., 1: 279–287.Google Scholar
  5. 5.
    Formal SB, Gemski P Jr, Baron LS, LaBrec EH, 1971, A chromosomal locus which controls the ability of Shigella flexneri to evoke keratoconjunctivitis, Infect Immun., 3:73–79.Google Scholar
  6. 6.
    Formal SB, LaBrec EH, Kent TH, Falkow S, 1965, Abortive intestinal infection with an Escherichia coli-Shigella flexneri hybrid strain, J Bacteriol., 89:1374–1382.Google Scholar
  7. 7.
    7„ LaBrec EH, Schneider H, Magnani TJ, Formal SB, 1964, Epithelial cell penetration as an essential step in the pathogenesis of bacillary dysentery, J Bacteriol., 88:1503–1518.Google Scholar
  8. 8.
    Lett MC, Sasakawa C, Okada N, Sakai T, Makino S, Yamada M, Komatsu K, Yoshikawa M, Identification of the virG protein and determination of the complete coding sequence: A plasmid-coded virulence gene of Shigella flexneri, J Bacteriol, in press.Google Scholar
  9. 9.
    Luria SE, Burrous JW, 1957, Hybridization between Escherichia coli and Shigella, J Bacteriol., 74:461–476.CrossRefGoogle Scholar
  10. 10.
    Makino S, Sasakawa C, Kamata K, Kurata T, Yoshikawa M, 1986, A genetic determinant required for continuous reinfection of adjacent cells on large plasmid in S. flexneri 2a, Cell, 46:551–555.CrossRefGoogle Scholar
  11. 11.
    Maurelli AT, Baudry B, d’Hauteville H, Hale TL, Sansonetti PJ, 1985, Cloning of plasmid DNA sequences involved in invasion of HeLa Cells by Shigella flexneri, Infect Immun., 49:164–171.Google Scholar
  12. 12.
    Maurelli AT, Blackmon B, Curtiss R III, 1984, Loss of pigmentation in Shigella flexneri 2a is correlated with loss of virulence and virulence- associated plasmid, Infect Immun., 43:397–401.Google Scholar
  13. 13.
    Murayama SY, Sakai T, Makino S, Kurata T, Sasakawa C, Yoshikawa M, 1986, The use of mice in the Sereny test as a virulence assay of shigellae and enteroinvasive Escherichia coli, Infect Immun., 51:696–698.Google Scholar
  14. 14.
    Oaks EV, Wingfield ME, Formal SB, 1985, Plaque formation by virulent Shigella flexneri, Infect Immun., 48:124–129.Google Scholar
  15. 15.
    Ogawa H, Nakamura A, Nakaya R, 1968, Cinemicrographic study of tissue cell cultures infected with Shigella flexneri, Japan J Med Sci Biol., 21:259–273.Google Scholar
  16. 16.
    Sakai T, Sasakawa C, Makino S, Kamata K, Yoshikawa M, 1986, Molecular cloning of a genetic determinant for Congo red binding ability which is essential for the virulence of Shigella flexneri, Infect Immun., 54: 395–402.Google Scholar
  17. 17.
    Sakai T, Sasakawa C, Yoshikawa M, 1986, DNA sequence and product analysis of the virF locus responsible for Congo red binding and cell invasion in Shigella flexneri 2a, Infect Immun., 54:395–402.Google Scholar
  18. 18.
    Sakai T, Sasakawa C, Yoshikawa M, 1988, Expression of four virulence antigens of Shigella flexneri is positively regulated at the transcriptional level by the 30 kilodalton virF protein, Molec Microbiol., 2: 589–597.CrossRefGoogle Scholar
  19. 19.
    Sansonetti PJ, Hale TL, Dammin GJ, Kapfer C, Collins HH Jr, Formal SB, 1983, Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri, Infect Immun., 39:1392–1402.Google Scholar
  20. 20.
    Sansonetti PJ, Kopecko DJ, Formal SB, 1982, Involvement of a plasmid in the invasive ability of Shigella flexneri, Infect Immun., 35:852–860.Google Scholar
  21. 21.
    Sansonetti PJ, Ryter A, Clerc P, Maurelli AT, Mounier J, 1986, Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and piasmid-mediated contact hemolysis, Infect Immun, 51:461–469.Google Scholar
  22. 22.
    Sasakawa C, Kamata K, Sakai T, Makino S, Yamada M, Okada N, Yoshikawa M, 1988, Virulence - associated genetic regions comprising 31 kilobases of the 230-kilobase plasmid in Shigella flexneri 2a, J Bacteriol., 170: 2480–2484.Google Scholar
  23. 23.
    Sasakawa C, Kamata K, Sakai T, Murayama SY, Makino S, Yoshikawa M, 1986, Molecular alteration of the 140-megadalton plasmid associated with loss of virulence and Congo red binding activity in Shigella flexneri, Infect Immun., 51:470–475.Google Scholar
  24. 24.
    Sasakawa C, Makino S, Kamata K, Yoshikawa M, 1986, 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., 54:32–36.Google Scholar
  25. 25.
    Sasakawa C, Yoshikawa M, 1987, A series of Tn5 variants with various drug-resistance markers and suicide vector for transposon mutagenesis, Gene, 56:283–288.CrossRefGoogle Scholar
  26. 26.
    Sereny B, 1955, Experimental shigella keratoconjunctivitis, Acta Microbiol Acad Sci Hung., 2:293–296.Google Scholar
  27. 27.
    Watanabe H, Nakamura A, 1985, Large plasmids associated with virulence in Shigella species have a common function necessary for epithelial penetration, Infect Immun., 48:260–262.Google Scholar
  28. 38.
    Watanabe H, Nakamura A, 1986, Identification of Shigella sonnei form I plasmid genes necessary for cell invasion and their conservation among Shigella species and enteroinvasive Escherichia coli, Infect Immun., 53:352–358.Google Scholar
  29. 29.
    Yamada M, Sasakawa C, Okada N, Makino S, Yoshikawa M, Molecular cloning and characterization of chromosomal virulence region KcpA of Shigella flexneri, Molec Microbiol., in press.Google Scholar

Copyright information

© Plenum Press, New York 1990

Authors and Affiliations

  • Masanosuke Yoshikawa
    • 1
  • Chihiro Sasakawa
    • 1
  • Takashi Sakai
    • 1
  • Masatoshi Yamada
    • 1
  • Nobuhiko Okada
    • 1
  • Sou-ichi Makino
    • 1
  1. 1.Department of Bacteriology The Institute of Medical ScienceThe University of TokyoTokyoJapan

Personalised recommendations