DOI: 10.1111/j.1365-2958.1996.tb02656.x. Effect of O side‐chain length and composition on the virulence of <i>Shigella flexneri</i> 2a

Effect of O side‐chain length and composition on the virulence of Shigella flexneri 2a

10.1111/j.1365-2958.1996.tb02656.x

Crossref journal-article

Wiley

Molecular Microbiology (311)

Abstract

SummaryIcsA of Shigella flexneri is required for intercellular spread and is located in the outer membrane at one pole of the bacterium, where it catalyses the polymerization of host‐cell actin. The formation of the actin tail provides the force to move the bacterium in a unidirectional manner through the host‐cell cytoplasm. We have previously demonstrated that rough lipopolysaccharide (LPS) mutants of S. flexneri 2a are avirulent and cannot form plaques in tissue‐culture monolayers. This inability to form plaques is associated with non‐polar localization of IcsA and loss of host‐cell membrane‐protrusion formation ('fireworks'). To define the minimal LPS structure required for fireworks formation, we constructed a strain of S. flexneri (BS497) that contains a mutation in rfc, encoding the O side‐chain polymerase, and a strain, BS520, that possesses a defective O side‐chain ligase due to a mutation in rfaL. BS497 produces a LPS that consists of a core with one repeat unit of the O side‐chain, while BS520 produces a LPS consisting of a complete core with no O side‐chain. BS497 remained invasive but did not form fireworks or plaques in tissue‐culture monolayers and was negative in the Serény test. BS520 was invasive, generated reduced numbers of short fireworks, and made tiny plaques, but it was negative in the Serény test. Analysis of BS497 with anti‐lcsA antibody demonstrated that IcsA was distributed over the entire cell surface. The distribution of IcsA on the surface of BS520 was predominantly unipolar, with some trail‐back of IcsA label along the sides of the bacterium. A similar pattern was seen when infected monolayers were stained for polymerized actin. These results suggest that both the presence and the length of the O side‐chain are important in the proper localization or maintenance of IcsA at the pole which subsequently affects the ability to form actin tails and produce fireworks. This reduced ability to form actin tails and fireworks results in a decreased ability of Shigella to move into adjacent host cells. To determine if the sugar composition of the O side‐chain is important in the ability to form fireworks, the rfb region of S. flexneri2a was replaced with the rfb region from Escherichia coll serotype O8 or O25. Both hybrids were invasive, formed plaques, and gave positive Serény reactions. These results suggest that, unlike LPS length, the sugar composition of the O side‐chain is not a critical requirement for the proper localization of IcsA and efficient intercellular movement.

Bibliography

Sandlin, R. C., Goldberg, M. B., & Maurelli, A. T. (1996). Effect of O sideâchain length and composition on the virulence of Shigella flexneri 2a. Molecular Microbiology, 22(1), 63â73. Portico.

Authors 3

Robin C. Sandlin (first)
Marcia B. Goldberg (additional)
Anthony T. Maurelli (additional)

References 33 Referenced 75

10.1073/pnas.86.10.3867
10.1111/j.1365-2958.1994.tb00434.x
10.1128/iai.6.2.104-111.1972 / Infect Immun / Virulence of Shigella flexneri hybrids expressing Escherichia coli somatic antigens by Gemski P. (1972)
10.1073/pnas.92.14.6572
10.1128/jb.175.8.2189-2196.1993
10.1128/iai.62.12.5664-5668.1994 / Infect Immun / Regulation of surface presentation of IcsA, a Shigella protein essential to intracellular movement and spread, is growth phase dependent by Goldberg M.B. (1994)
10.1128/MMBR.55.2.206-224.1991
10.1128/JB.154.1.269-277.1983
10.1128/iai.57.10.2963-2970.1989 / Infect Immun / Identification of Shigella invasion genes by isolation of temperature‐regulated inv:: lacZ operon fusions by Hromockyj A.E. (1989)
10.1016/0008-6215(83)88336-0
10.1111/j.1365-2958.1993.tb01700.x
10.1128/jb.174.22.7297-7307.1992
10.1111/j.1365-2958.1995.mmi_18030413.x
10.1128/JB.88.5.1503-1518.1964
10.1016/0962-8924(96)81001-4
10.1093/clinids/13.Supplement_4.S279
10.1128/jb.175.22.7125-7129.1993
10.1128/jb.163.2.756-762.1985 / J Bacteriol / Genetic location of genes encoding enterobacterial common antigen by Meier U. (1985)
10.1016/S0021-9258(18)77373-0 / J Biol Chem / Biosynthesis of enterobacterial common antigen in Escherichia coli: biochemical characterization of Tn70 insertion mutants defective in enterobacterial common antigen synthesis by Meier‐Dieter U. (1990)
10.1016/S0021-9258(18)48347-0 / J Biol Chem / Nucleotide sequence of the Escherichia coli rfe gene involved in the synthesis of enterobacterial common antigen by Meier‐Dieter U. (1992)
{'key': 'e_1_2_1_22_1', 'volume-title': 'Experiments in Molecular Genetics', 'author': 'Miller J.H.', 'year': '1972'} / Experiments in Molecular Genetics by Miller J.H. (1972)
10.1128/jb.170.6.2575-2583.1988
10.1128/jb.176.3.733-747.1994 / J Bacteriol / Characterization of the rfe region of Shigella flexneri by Morona R. (1994)
10.1128/IAI.48.1.124-129.1985
{'key': 'e_1_2_1_26_1', 'first-page': '648', 'volume-title': 'Escherichia coli and Salmonella typhimurium; Cellular and Molecular Biology', 'author': 'Rick P.D.', 'year': '1987'} / Escherichia coli and Salmonella typhimurium; Cellular and Molecular Biology by Rick P.D. (1987)
10.1128/jb.176.10.2877-2884.1994
10.1128/IAI.63.1.229-237.1995
10.1016/0264-410X(91)90128-S
{'key': 'e_1_2_1_30_1', 'first-page': '367', 'article-title': 'Experimental keratoconjunctivitis shigellosa', 'volume': '4', 'author': 'Serény B.', 'year': '1957', 'journal-title': 'Acta Microbiol Acad Sci Hung'} / Acta Microbiol Acad Sci Hung / Experimental keratoconjunctivitis shigellosa by Serény B. (1957)
10.1099/00222615-23-4-289
10.1111/j.1365-2958.1995.mmi_17050945.x
10.1016/0003-2697(82)90673-X
10.1016/0378-1135(92)90102-Y

Dates

Type	When
Created	18 years, 10 months ago (Oct. 6, 2006, 6:28 a.m.)
Deposited	1 year, 10 months ago (Oct. 26, 2023, 5:02 p.m.)
Indexed	1 month, 1 week ago (July 14, 2025, 11:33 p.m.)
Issued	28 years, 10 months ago (Oct. 1, 1996)
Published	28 years, 10 months ago (Oct. 1, 1996)
Published Online	18 years, 10 months ago (Oct. 6, 2006)
Published Print	28 years, 10 months ago (Oct. 1, 1996)

Funders 0

None

BibTeX

@article{Sandlin_1996, title={Effect of O side‐chain length and composition on the virulence of Shigella flexneri 2a}, volume={22}, ISSN={1365-2958}, url={http://dx.doi.org/10.1111/j.1365-2958.1996.tb02656.x}, DOI={10.1111/j.1365-2958.1996.tb02656.x}, number={1}, journal={Molecular Microbiology}, publisher={Wiley}, author={Sandlin, Robin C. and Goldberg, Marcia B. and Maurelli, Anthony T.}, year={1996}, month=oct, pages={63–73} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        15
      ]
    ],
    "date-time": "2025-07-15T03:33:33Z",
    "timestamp": 1752550413352
  },
  "reference-count": 33,
  "publisher": "Wiley",
  "issue": "1",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2006,
            10,
            6
          ]
        ],
        "date-time": "2006-10-06T00:00:00Z",
        "timestamp": 1160092800000
      },
      "content-version": "vor",
      "delay-in-days": 3657,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1996,
        10
      ]
    ]
  },
  "abstract": "<jats:title>Summary</jats:title><jats:p>IcsA of <jats:italic>Shigella flexneri</jats:italic> is required for intercellular spread and is located in the outer membrane at one pole of the bacterium, where it catalyses the polymerization of host\u2010cell actin. The formation of the actin tail provides the force to move the bacterium in a unidirectional manner through the host\u2010cell cytoplasm. We have previously demonstrated that rough lipopolysaccharide (LPS) mutants of S. <jats:italic>flexneri</jats:italic> 2a are avirulent and cannot form plaques in tissue\u2010culture monolayers. This inability to form plaques is associated with non\u2010polar localization of IcsA and loss of host\u2010cell membrane\u2010protrusion formation ('fireworks'). To define the minimal LPS structure required for fireworks formation, we constructed a strain of S. <jats:italic>flexneri</jats:italic> (BS497) that contains a mutation in <jats:italic>rfc</jats:italic>, encoding the O side\u2010chain polymerase, and a strain, BS520, that possesses a defective O side\u2010chain ligase due to a mutation in <jats:italic>rfaL.</jats:italic> BS497 produces a LPS that consists of a core with one repeat unit of the O side\u2010chain, while BS520 produces a LPS consisting of a complete core with no O side\u2010chain. BS497 remained invasive but did not form fireworks or plaques in tissue\u2010culture monolayers and was negative in the Ser\u00e9ny test. BS520 was invasive, generated reduced numbers of short fireworks, and made tiny plaques, but it was negative in the Ser\u00e9ny test. Analysis of BS497 with anti\u2010lcsA antibody demonstrated that IcsA was distributed over the entire cell surface. The distribution of IcsA on the surface of BS520 was predominantly unipolar, with some trail\u2010back of IcsA label along the sides of the bacterium. A similar pattern was seen when infected monolayers were stained for polymerized actin. These results suggest that both the presence and the length of the O side\u2010chain are important in the proper localization or maintenance of IcsA at the pole which subsequently affects the ability to form actin tails and produce fireworks. This reduced ability to form actin tails and fireworks results in a decreased ability of <jats:italic>Shigella</jats:italic> to move into adjacent host cells. To determine if the sugar composition of the O side\u2010chain is important in the ability to form fireworks, the <jats:italic>rfb</jats:italic> region of S. <jats:italic>flexneri2a</jats:italic> was replaced with the <jats:italic>rfb</jats:italic> region from <jats:italic>Escherichia coll</jats:italic> serotype O8 or O25. Both hybrids were invasive, formed plaques, and gave positive Ser\u00e9ny reactions. These results suggest that, unlike LPS length, the sugar composition of the O side\u2010chain is not a critical requirement for the proper localization of IcsA and efficient intercellular movement.</jats:p>",
  "DOI": "10.1111/j.1365-2958.1996.tb02656.x",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2006,
        10,
        6
      ]
    ],
    "date-time": "2006-10-06T10:28:10Z",
    "timestamp": 1160130490000
  },
  "page": "63-73",
  "source": "Crossref",
  "is-referenced-by-count": 75,
  "title": "Effect of O side\u2010chain length and composition on the virulence of <i>Shigella flexneri</i> 2a",
  "prefix": "10.1111",
  "volume": "22",
  "author": [
    {
      "given": "Robin C.",
      "family": "Sandlin",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Marcia B.",
      "family": "Goldberg",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Anthony T.",
      "family": "Maurelli",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2006,
        10,
        6
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_1_2_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.86.10.3867"
    },
    {
      "key": "e_1_2_1_3_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.1994.tb00434.x"
    },
    {
      "key": "e_1_2_1_4_1",
      "doi-asserted-by": "crossref",
      "first-page": "104",
      "DOI": "10.1128/iai.6.2.104-111.1972",
      "article-title": "Virulence of Shigella flexneri hybrids expressing Escherichia coli somatic antigens",
      "volume": "6",
      "author": "Gemski P.",
      "year": "1972",
      "journal-title": "Infect Immun"
    },
    {
      "key": "e_1_2_1_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.92.14.6572"
    },
    {
      "key": "e_1_2_1_6_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.175.8.2189-2196.1993"
    },
    {
      "key": "e_1_2_1_7_1",
      "doi-asserted-by": "crossref",
      "first-page": "5664",
      "DOI": "10.1128/iai.62.12.5664-5668.1994",
      "article-title": "Regulation of surface presentation of IcsA, a Shigella protein essential to intracellular movement and spread, is growth phase dependent",
      "volume": "62",
      "author": "Goldberg M.B.",
      "year": "1994",
      "journal-title": "Infect Immun"
    },
    {
      "key": "e_1_2_1_8_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MMBR.55.2.206-224.1991"
    },
    {
      "key": "e_1_2_1_9_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.154.1.269-277.1983"
    },
    {
      "key": "e_1_2_1_10_1",
      "doi-asserted-by": "crossref",
      "first-page": "2963",
      "DOI": "10.1128/iai.57.10.2963-2970.1989",
      "article-title": "Identification of Shigella invasion genes by isolation of temperature\u2010regulated inv:: lacZ operon fusions",
      "volume": "57",
      "author": "Hromockyj A.E.",
      "year": "1989",
      "journal-title": "Infect Immun"
    },
    {
      "key": "e_1_2_1_11_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0008-6215(83)88336-0"
    },
    {
      "key": "e_1_2_1_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.1993.tb01700.x"
    },
    {
      "key": "e_1_2_1_13_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.174.22.7297-7307.1992"
    },
    {
      "key": "e_1_2_1_14_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.1995.mmi_18030413.x"
    },
    {
      "key": "e_1_2_1_15_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.88.5.1503-1518.1964"
    },
    {
      "key": "e_1_2_1_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0962-8924(96)81001-4"
    },
    {
      "key": "e_1_2_1_17_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/clinids/13.Supplement_4.S279"
    },
    {
      "key": "e_1_2_1_18_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.175.22.7125-7129.1993"
    },
    {
      "key": "e_1_2_1_19_1",
      "doi-asserted-by": "crossref",
      "first-page": "756",
      "DOI": "10.1128/jb.163.2.756-762.1985",
      "article-title": "Genetic location of genes encoding enterobacterial common antigen",
      "volume": "163",
      "author": "Meier U.",
      "year": "1985",
      "journal-title": "J Bacteriol"
    },
    {
      "key": "e_1_2_1_20_1",
      "doi-asserted-by": "crossref",
      "first-page": "13490",
      "DOI": "10.1016/S0021-9258(18)77373-0",
      "article-title": "Biosynthesis of enterobacterial common antigen in Escherichia coli: biochemical characterization of Tn70 insertion mutants defective in enterobacterial common antigen synthesis",
      "volume": "265",
      "author": "Meier\u2010Dieter U.",
      "year": "1990",
      "journal-title": "J Biol Chem"
    },
    {
      "key": "e_1_2_1_21_1",
      "doi-asserted-by": "crossref",
      "first-page": "746",
      "DOI": "10.1016/S0021-9258(18)48347-0",
      "article-title": "Nucleotide sequence of the Escherichia coli rfe gene involved in the synthesis of enterobacterial common antigen",
      "volume": "267",
      "author": "Meier\u2010Dieter U.",
      "year": "1992",
      "journal-title": "J Biol Chem"
    },
    {
      "key": "e_1_2_1_22_1",
      "volume-title": "Experiments in Molecular Genetics",
      "author": "Miller J.H.",
      "year": "1972"
    },
    {
      "key": "e_1_2_1_23_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.170.6.2575-2583.1988"
    },
    {
      "key": "e_1_2_1_24_1",
      "doi-asserted-by": "crossref",
      "first-page": "733",
      "DOI": "10.1128/jb.176.3.733-747.1994",
      "article-title": "Characterization of the rfe region of Shigella flexneri",
      "volume": "176",
      "author": "Morona R.",
      "year": "1994",
      "journal-title": "J Bacteriol"
    },
    {
      "key": "e_1_2_1_25_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/IAI.48.1.124-129.1985"
    },
    {
      "key": "e_1_2_1_26_1",
      "first-page": "648",
      "volume-title": "Escherichia coli and Salmonella typhimurium; Cellular and Molecular Biology",
      "author": "Rick P.D.",
      "year": "1987"
    },
    {
      "key": "e_1_2_1_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.176.10.2877-2884.1994"
    },
    {
      "key": "e_1_2_1_28_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/IAI.63.1.229-237.1995"
    },
    {
      "key": "e_1_2_1_29_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0264-410X(91)90128-S"
    },
    {
      "key": "e_1_2_1_30_1",
      "first-page": "367",
      "article-title": "Experimental keratoconjunctivitis shigellosa",
      "volume": "4",
      "author": "Ser\u00e9ny B.",
      "year": "1957",
      "journal-title": "Acta Microbiol Acad Sci Hung"
    },
    {
      "key": "e_1_2_1_31_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/00222615-23-4-289"
    },
    {
      "key": "e_1_2_1_32_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.1995.mmi_17050945.x"
    },
    {
      "key": "e_1_2_1_33_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-2697(82)90673-X"
    },
    {
      "key": "e_1_2_1_34_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0378-1135(92)90102-Y"
    }
  ],
  "container-title": "Molecular Microbiology",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1365-2958.1996.tb02656.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2958.1996.tb02656.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        26
      ]
    ],
    "date-time": "2023-10-26T21:02:27Z",
    "timestamp": 1698354147000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://onlinelibrary.wiley.com/doi/10.1111/j.1365-2958.1996.tb02656.x"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1996,
        10
      ]
    ]
  },
  "references-count": 33,
  "journal-issue": {
    "issue": "1",
    "published-print": {
      "date-parts": [
        [
          1996,
          10
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1111/j.1365-2958.1996.tb02656.x"
  ],
  "URL": "http://dx.doi.org/10.1111/j.1365-2958.1996.tb02656.x",
  "relation": {},
  "ISSN": [
    "0950-382X",
    "1365-2958"
  ],
  "subject": [],
  "container-title-short": "Molecular Microbiology",
  "published": {
    "date-parts": [
      [
        1996,
        10
      ]
    ]
  }
}