DOI: 10.1126/science.274.5288.780. A Role for Phosphoinositide 3-Kinase in Bacterial Invasion

A Role for Phosphoinositide 3-Kinase in Bacterial Invasion

10.1126/science.274.5288.780

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Listeria monocytogenes is a bacterial pathogen that invades cultured nonphagocytic cells. Inhibitors and a dominant negative mutation were used to demonstrate that efficient entry requires the phosphoinositide (PI) 3-kinase p85α-p110. Infection with L. monocytogenes caused rapid increases in cellular amounts of PI(3,4)P 2 and PI(3,4,5)P 3 , indicating that invading bacteria stimulated PI 3-kinase activity. This stimulation required the bacterial protein InlB, host cell tyrosine phosphorylation, and association of p85α with one or more tyrosine-phosphorylated proteins. This role for PI 3-kinase in bacterial entry may have parallels in some endocytic events.

Bibliography

Ireton, K., Payrastre, B., Chap, H., Ogawa, W., Sakaue, H., Kasuga, M., & Cossart, P. (1996). A Role for Phosphoinositide 3-Kinase in Bacterial Invasion. Science, 274(5288), 780â782.

Authors 7

Keith Ireton (first)
Bernard Payrastre (additional)
Hugues Chap (additional)
Wataru Ogawa (additional)
Hiroshi Sakaue (additional)
Masato Kasuga (additional)
Pascale Cossart (additional)

References 21 Referenced 270

Dramsi S., Lebrun M., Cossart P., Curr. Top. Microbiol. Immunol. 209611996. / Curr. Top. Microbiol. Immunol. by Dramsi S. (1996)
10.1016/S0092-8674(00)81070-3
Tang P., Rosenshine I., Finlay B. B., Mol. Biol. Cell 54551994; P. Velge et al.,Microb. Pathog.17, 37 (1994). (10.1091/mbc.5.4.455) / Mol. Biol. Cell by Tang P. (1994)
Carpenter C. L., Cantley L. C., Curr. Opin. Cell Biol. 81531996. (10.1016/S0955-0674(96)80060-3) / Curr. Opin. Cell Biol. by Carpenter C. L. (1996)
Shepherd P. R., Reaves B. J., Davidson H. W., Trends Cell Biol. 6921996. (10.1016/0962-8924(96)80998-6) / Trends Cell Biol. by Shepherd P. R. (1996)
Ui M., Okada T., Hazeki K., Hazeki O., Trends Biochem. Sci. 203031995. (10.1016/S0968-0004(00)89056-8) / Trends Biochem. Sci. by Ui M. (1995)
10.1016/S0021-9258(17)37680-9
Rameh L. E., Chen C. S., Cantley L. C., Cell 838211996. (10.1016/0092-8674(95)90195-7) / Cell by Rameh L. E. (1996)
Ireton K. Cossart P. unpublished data.
Hara K., et al., Proc. Natl. Acad. Sci. U.S.A. 9174151994. (10.1073/pnas.91.16.7415) / Proc. Natl. Acad. Sci. U.S.A. by Hara K. (1994)
Kasuga M. Ogawa W. Sakaue H. in preparation.
Percent invasion values are the means ± SD of four experiments. Construction of CHO-Δp85α cells will be described elsewhere (11). CHO and CHO-Δp85α cells were grown as described (10) and used between passages 4 to 8 and 5 to 8 respectively. Hygromycin (150 ng/ml) was included in the medium for routine growth of CHO-Δp85α cells except when seeded for invasion tests. Invasion assays were performed on 2-day-old monolayers in 24-well tissue culture plates (about 4 × 104 cells per well). Expression of Δp85α did not affect bacterial adhesion or cell viability as determined by Trypan blue staining.
10.1111/j.1365-2958.1995.tb02297.x
10.1016/0092-8674(91)90009-N
Hartwig J. H., et al., ibid. 826431995. / ibid. by Hartwig J. H. (1995)
Guinebault C., et al., J. Cell Biol. 1298311995. (10.1083/jcb.129.3.831) / J. Cell Biol. by Guinebault C. (1995)
Gout I., et al., Cell 75251993; A. Ando et al., EMBO J.13, 3033 (1994). (10.1016/S0092-8674(05)80081-9) / Cell by Gout I. (1993)
Vero cells were grown in Dulbecco's modified Eagle's medium (DMEM) with fetal calf serum (10%) (Gibco) and Non-Essential Amino Acids (Gibco). Caco-2 cells were grown as described (13) and used between passages 78 and 86. Subconfluent monolayers of cells for invasion tests were obtained by seeding 2 × 104 Vero or 4 × 104 Caco-2 cells per well in 24-well tissue culture plates and incubating at 37°C in 10% C02 for 2 days before the test. Strain EGD (13) was prepared for invasion tests as described (2) and a multiplicity of infection (MOI) of about 100 bacteria per cell was used. Vero or Caco-2 cells were treated with wortmannin (Sigma or Biomol) or LY294002 (Biomol) 20 min before infection as well as during the 60-min infection. Inhibitors were added from 1000× solutions in DMSO and therefore DMSO was added at a final concentration of 0.1% in the control (0 concentration). At the concentrations used the inhibitors had no effect on cell or bacterial viability as assayed by the [3-(4 5-dimethylthiazol-2-yl)-2 5-diphenyl-tetrazolium bromide (MTT)] test (19) or titering of viable bacteria respectively. The inhibitors also did not affect bacterial adhesion to Vero or Caco-2 cells.
Hansen M. B., Nielsen S. E., Berg K., J. Immunol. Methods 1192031989. (10.1016/0022-1759(89)90397-9) / J. Immunol. Methods by Hansen M. B. (1989)
Similar increases in labeled PI(3 4)P2 and PI(3 4 5)P3 were observed when a smaller MOI was used (100:1) except that these increases were slightly smaller and apparent 15 to 30 min after infection.
Supported by grants from the Ministere de l'Agriculture et la Peche (R94-35) DRET (93-109) INSERM (CRE93013) the Ministere de la Recherche (ACCSV6) and the Pasteur Institute. K.I. is a recipient of a fellowship from the Jane Coffin Childs Memorial Fund for Medical Research. We thank P. Boquet for suggesting wortmannin and Jerôme Mengaud for critically reading the manuscript.

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 5:45 a.m.)
Deposited	1 year ago (Aug. 7, 2024, 7:27 a.m.)
Indexed	1 month, 3 weeks ago (July 11, 2025, 6:26 a.m.)
Issued	28 years, 10 months ago (Nov. 1, 1996)
Published	28 years, 10 months ago (Nov. 1, 1996)
Published Print	28 years, 10 months ago (Nov. 1, 1996)

Funders 0

None

BibTeX

@article{Ireton_1996, title={A Role for Phosphoinositide 3-Kinase in Bacterial Invasion}, volume={274}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.274.5288.780}, DOI={10.1126/science.274.5288.780}, number={5288}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Ireton, Keith and Payrastre, Bernard and Chap, Hugues and Ogawa, Wataru and Sakaue, Hiroshi and Kasuga, Masato and Cossart, Pascale}, year={1996}, month=nov, pages={780–782} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        11
      ]
    ],
    "date-time": "2025-07-11T10:26:51Z",
    "timestamp": 1752229611546
  },
  "reference-count": 21,
  "publisher": "American Association for the Advancement of Science (AAAS)",
  "issue": "5288",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1996,
        11
      ]
    ]
  },
  "abstract": "<jats:p>\n            <jats:italic>Listeria monocytogenes</jats:italic>\n            is a bacterial pathogen that invades cultured nonphagocytic cells. Inhibitors and a dominant negative mutation were used to demonstrate that efficient entry requires the phosphoinositide (PI) 3-kinase p85\u03b1-p110. Infection with\n            <jats:italic>L. monocytogenes</jats:italic>\n            caused rapid increases in cellular amounts of PI(3,4)P\n            <jats:sub>2</jats:sub>\n            and PI(3,4,5)P\n            <jats:sub>3</jats:sub>\n            , indicating that invading bacteria stimulated PI 3-kinase activity. This stimulation required the bacterial protein InlB, host cell tyrosine phosphorylation, and association of p85\u03b1 with one or more tyrosine-phosphorylated proteins. This role for PI 3-kinase in bacterial entry may have parallels in some endocytic events.\n          </jats:p>",
  "DOI": "10.1126/science.274.5288.780",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        27
      ]
    ],
    "date-time": "2002-07-27T09:45:50Z",
    "timestamp": 1027763150000
  },
  "page": "780-782",
  "source": "Crossref",
  "is-referenced-by-count": 270,
  "title": "A Role for Phosphoinositide 3-Kinase in Bacterial Invasion",
  "prefix": "10.1126",
  "volume": "274",
  "author": [
    {
      "given": "Keith",
      "family": "Ireton",
      "sequence": "first",
      "affiliation": [
        {
          "name": "K. Ireton and P. Cossart, Unit\u00e9 des Interactions Bact\u00e9ries-Cellules, Institut Pasteur, 75724 Paris Cedex 15, France."
        }
      ]
    },
    {
      "given": "Bernard",
      "family": "Payrastre",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "B. Payrastre and H. Chap, INSERM Unit\u00e9 326, H\u00f4pital Purpan, 31059 Toulouse, France."
        }
      ]
    },
    {
      "given": "Hugues",
      "family": "Chap",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "B. Payrastre and H. Chap, INSERM Unit\u00e9 326, H\u00f4pital Purpan, 31059 Toulouse, France."
        }
      ]
    },
    {
      "given": "Wataru",
      "family": "Ogawa",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "W. Ogawa, H. Sakaue, M. Kasuga, Second Department of Internal Medicine, Kobe University School of Medicine, Chuo-ku, Kobe 650, Japan."
        }
      ]
    },
    {
      "given": "Hiroshi",
      "family": "Sakaue",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "W. Ogawa, H. Sakaue, M. Kasuga, Second Department of Internal Medicine, Kobe University School of Medicine, Chuo-ku, Kobe 650, Japan."
        }
      ]
    },
    {
      "given": "Masato",
      "family": "Kasuga",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "W. Ogawa, H. Sakaue, M. Kasuga, Second Department of Internal Medicine, Kobe University School of Medicine, Chuo-ku, Kobe 650, Japan."
        }
      ]
    },
    {
      "given": "Pascale",
      "family": "Cossart",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "K. Ireton and P. Cossart, Unit\u00e9 des Interactions Bact\u00e9ries-Cellules, Institut Pasteur, 75724 Paris Cedex 15, France."
        }
      ]
    }
  ],
  "member": "221",
  "reference": [
    {
      "key": "e_1_3_1_2_2",
      "first-page": "61",
      "volume": "209",
      "author": "Dramsi S.",
      "year": "1996",
      "unstructured": "Dramsi S., Lebrun M., Cossart P., Curr. Top. Microbiol. Immunol. 209611996.",
      "journal-title": "Curr. Top. Microbiol. Immunol."
    },
    {
      "key": "e_1_3_1_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0092-8674(00)81070-3"
    },
    {
      "key": "e_1_3_1_4_2",
      "doi-asserted-by": "crossref",
      "first-page": "455",
      "DOI": "10.1091/mbc.5.4.455",
      "volume": "5",
      "author": "Tang P.",
      "year": "1994",
      "unstructured": "Tang P., Rosenshine I., Finlay B. B., Mol. Biol. Cell 54551994; P. Velge et al.,Microb. Pathog.17, 37 (1994).",
      "journal-title": "Mol. Biol. Cell"
    },
    {
      "key": "e_1_3_1_5_2",
      "doi-asserted-by": "crossref",
      "first-page": "153",
      "DOI": "10.1016/S0955-0674(96)80060-3",
      "volume": "8",
      "author": "Carpenter C. L.",
      "year": "1996",
      "unstructured": "Carpenter C. L., Cantley L. C., Curr. Opin. Cell Biol. 81531996.",
      "journal-title": "Curr. Opin. Cell Biol."
    },
    {
      "key": "e_1_3_1_6_2",
      "doi-asserted-by": "crossref",
      "first-page": "92",
      "DOI": "10.1016/0962-8924(96)80998-6",
      "volume": "6",
      "author": "Shepherd P. R.",
      "year": "1996",
      "unstructured": "Shepherd P. R., Reaves B. J., Davidson H. W., Trends Cell Biol. 6921996.",
      "journal-title": "Trends Cell Biol."
    },
    {
      "key": "e_1_3_1_7_2",
      "doi-asserted-by": "crossref",
      "first-page": "303",
      "DOI": "10.1016/S0968-0004(00)89056-8",
      "volume": "20",
      "author": "Ui M.",
      "year": "1995",
      "unstructured": "Ui M., Okada T., Hazeki K., Hazeki O., Trends Biochem. Sci. 203031995.",
      "journal-title": "Trends Biochem. Sci."
    },
    {
      "key": "e_1_3_1_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(17)37680-9"
    },
    {
      "key": "e_1_3_1_9_2",
      "doi-asserted-by": "crossref",
      "first-page": "821",
      "DOI": "10.1016/0092-8674(95)90195-7",
      "volume": "83",
      "author": "Rameh L. E.",
      "year": "1996",
      "unstructured": "Rameh L. E., Chen C. S., Cantley L. C., Cell 838211996.",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_10_2",
      "unstructured": "Ireton K. Cossart P. unpublished data."
    },
    {
      "key": "e_1_3_1_11_2",
      "doi-asserted-by": "crossref",
      "first-page": "7415",
      "DOI": "10.1073/pnas.91.16.7415",
      "volume": "91",
      "author": "Hara K.",
      "year": "1994",
      "unstructured": "Hara K., et al., Proc. Natl. Acad. Sci. U.S.A. 9174151994.",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_12_2",
      "unstructured": "Kasuga M. Ogawa W. Sakaue H. in preparation."
    },
    {
      "key": "e_1_3_1_13_2",
      "unstructured": "Percent invasion values are the means \u00b1 SD of four experiments. Construction of CHO-\u0394p85\u03b1 cells will be described elsewhere (11). CHO and CHO-\u0394p85\u03b1 cells were grown as described (10) and used between passages 4 to 8 and 5 to 8 respectively. Hygromycin (150 ng/ml) was included in the medium for routine growth of CHO-\u0394p85\u03b1 cells except when seeded for invasion tests. Invasion assays were performed on 2-day-old monolayers in 24-well tissue culture plates (about 4 \u00d7 104 cells per well). Expression of \u0394p85\u03b1 did not affect bacterial adhesion or cell viability as determined by Trypan blue staining."
    },
    {
      "key": "e_1_3_1_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.1995.tb02297.x"
    },
    {
      "key": "e_1_3_1_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(91)90009-N"
    },
    {
      "key": "e_1_3_1_16_2",
      "first-page": "643",
      "volume": "82",
      "author": "Hartwig J. H.",
      "year": "1995",
      "unstructured": "Hartwig J. H., et al., ibid. 826431995.",
      "journal-title": "ibid."
    },
    {
      "key": "e_1_3_1_17_2",
      "doi-asserted-by": "crossref",
      "first-page": "831",
      "DOI": "10.1083/jcb.129.3.831",
      "volume": "129",
      "author": "Guinebault C.",
      "year": "1995",
      "unstructured": "Guinebault C., et al., J. Cell Biol. 1298311995.",
      "journal-title": "J. Cell Biol."
    },
    {
      "key": "e_1_3_1_18_2",
      "doi-asserted-by": "crossref",
      "first-page": "25",
      "DOI": "10.1016/S0092-8674(05)80081-9",
      "volume": "75",
      "author": "Gout I.",
      "year": "1993",
      "unstructured": "Gout I., et al., Cell 75251993; A. Ando et al., EMBO J.13, 3033 (1994).",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_19_2",
      "unstructured": "Vero cells were grown in Dulbecco's modified Eagle's medium (DMEM) with fetal calf serum (10%) (Gibco) and Non-Essential Amino Acids (Gibco). Caco-2 cells were grown as described (13) and used between passages 78 and 86. Subconfluent monolayers of cells for invasion tests were obtained by seeding 2 \u00d7 104 Vero or 4 \u00d7 104 Caco-2 cells per well in 24-well tissue culture plates and incubating at 37\u00b0C in 10% C02 for 2 days before the test. Strain EGD (13) was prepared for invasion tests as described (2) and a multiplicity of infection (MOI) of about 100 bacteria per cell was used. Vero or Caco-2 cells were treated with wortmannin (Sigma or Biomol) or LY294002 (Biomol) 20 min before infection as well as during the 60-min infection. Inhibitors were added from 1000\u00d7 solutions in DMSO and therefore DMSO was added at a final concentration of 0.1% in the control (0 concentration). At the concentrations used the inhibitors had no effect on cell or bacterial viability as assayed by the [3-(4 5-dimethylthiazol-2-yl)-2 5-diphenyl-tetrazolium bromide (MTT)] test (19) or titering of viable bacteria respectively. The inhibitors also did not affect bacterial adhesion to Vero or Caco-2 cells."
    },
    {
      "key": "e_1_3_1_20_2",
      "doi-asserted-by": "crossref",
      "first-page": "203",
      "DOI": "10.1016/0022-1759(89)90397-9",
      "volume": "119",
      "author": "Hansen M. B.",
      "year": "1989",
      "unstructured": "Hansen M. B., Nielsen S. E., Berg K., J. Immunol. Methods 1192031989.",
      "journal-title": "J. Immunol. Methods"
    },
    {
      "key": "e_1_3_1_21_2",
      "unstructured": "Similar increases in labeled PI(3 4)P2 and PI(3 4 5)P3 were observed when a smaller MOI was used (100:1) except that these increases were slightly smaller and apparent 15 to 30 min after infection."
    },
    {
      "key": "e_1_3_1_22_2",
      "unstructured": "Supported by grants from the Ministere de l'Agriculture et la Peche (R94-35) DRET (93-109) INSERM (CRE93013) the Ministere de la Recherche (ACCSV6) and the Pasteur Institute. K.I. is a recipient of a fellowship from the Jane Coffin Childs Memorial Fund for Medical Research. We thank P. Boquet for suggesting wortmannin and Jer\u00f4me Mengaud for critically reading the manuscript."
    }
  ],
  "container-title": "Science",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://www.science.org/doi/pdf/10.1126/science.274.5288.780",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2024,
        8,
        7
      ]
    ],
    "date-time": "2024-08-07T11:27:43Z",
    "timestamp": 1723030063000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://www.science.org/doi/10.1126/science.274.5288.780"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1996,
        11
      ]
    ]
  },
  "references-count": 21,
  "journal-issue": {
    "issue": "5288",
    "published-print": {
      "date-parts": [
        [
          1996,
          11
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1126/science.274.5288.780"
  ],
  "URL": "http://dx.doi.org/10.1126/science.274.5288.780",
  "relation": {},
  "ISSN": [
    "0036-8075",
    "1095-9203"
  ],
  "subject": [],
  "container-title-short": "Science",
  "published": {
    "date-parts": [
      [
        1996,
        11
      ]
    ]
  }
}