DOI: 10.1046/j.1365-2958.2000.01771.x. Interactions among components of the <i>Salmonella</i> flagellar export apparatus and its substrates

Interactions among components of the Salmonella flagellar export apparatus and its substrates

10.1046/j.1365-2958.2000.01771.x

Crossref journal-article

Wiley

Molecular Microbiology (311)

Abstract

We have examined the cytoplasmic components (FliH, FliI and FliJ) of the type III flagellar protein export apparatus, plus the cytoplasmic domains (FlhAC and FlhBC) of two of its six membrane components. FliH, FlhAC and FliJ, when overproduced, caused inhibition of motility of wild‐type cells and inhibition of the export of substrates such as the hook protein FlgE. Co‐overproduction of FliH and FliI substantially relieved the inhibition caused by FliH, suggesting that it is excess free FliH that is inhibitory and that FliH and FliI form a complex. We purified His‐FLAG‐tagged versions of: (i) export components FliH, FliI, FliJ, FlhAC and FlhBC; (ii) rod/hook‐type export substrates FlgB (rod protein), FlgE (hook protein), FlgD (hook capping protein) and FliE (basal body protein); and (iii) filament‐type export substrates FlgK and FlgL (hook–filament junction proteins) and FliC (flagellin). We tested for protein–protein interactions by affinity blotting. In many cases, a given protein interacted with more than one other component, indicating that there are likely to be multiple dynamic interactions or interactions that involve more than two components. Interactions of FlhBC with rod/hook‐type substrates were strong, whereas those with filament‐type substrates were very weak; this may reflect the role of FlhB in substrate specificity switching. We propose a model for the flagellar export apparatus in which FlhA and FlhB and the other four integral membrane proteins of the apparatus form a complex at the base of the flagellar motor. A soluble complex of at least three proteins (FliH, FliI and FliJ) bind the protein to be exported and then interact with the complex at the motor to deliver the protein, which is then exported in an ATP‐dependent process mediated by FliI.

Bibliography

Minamino, T., & Macnab, R. M. (2000). Interactions among components of the Salmonella flagellar export apparatus and its substrates. Molecular Microbiology, 35(5), 1052â1064. Portico.

Authors 2

Tohru Minamino (first)
Robert M. Macnab (additional)

References 32 Referenced 178

10.1074/jbc.271.50.31981
10.1046/j.1365-2958.1997.6412010.x
10.1046/j.1365-2958.1999.01372.x
10.1016/S0022-2836(05)80266-9
10.1016/0022-2836(90)90365-S
10.1038/nbt1088-1204
10.1111/j.1365-2958.2005.04758.x
10.1126/science.8235660
10.1016/S0021-9258(17)36323-8 / J Biol Chem / The covalent structure of the phase‐1 flagellar filament protein of Salmonella typhimurium and its comparison with other flagellins by Joys T.M. (1985)
10.1006/jmbi.1996.0038
10.1128/jb.176.12.3598-3605.1994
10.1128/jb.176.24.7625-7629.1994
{'key': 'e_1_2_6_14_1', 'first-page': '123', 'volume-title': 'Escherichia coli and Salmonella: Cellular and Molecular Biology', 'author': 'Macnab R.M.', 'year': '1996'} / Escherichia coli and Salmonella: Cellular and Molecular Biology by Macnab R.M. (1996)
10.1128/JB.181.5.1388-1394.1999
10.1128/jb.176.24.7630-7637.1994
10.1046/j.1365-2958.1999.01597.x
10.1271/bbb.63.1301
10.1128/jb.174.7.2298-2304.1992 / J Bacteriol / Characterization of the fliE genes of Escherichia coli and Salmonella typhimurium and identification of the FliE protein as a component of the flagellar hook‐basal body complex by Müller V. (1992)
10.1046/j.1365-2958.1998.00998.x
10.1128/JB.181.18.5808-5813.1999
10.1128/JB.181.5.1555-1561.1999
10.1111/j.1365-2958.1992.tb01771.x
10.1128/jb.176.8.2272-2281.1994
10.1111/j.1742-4658.2006.05480.x
10.1128/JB.181.4.1059-1071.1999
10.1128/jb.179.17.5355-5365.1997
10.1006/jsbi.1998.4048
10.1128/jb.173.11.3564-3572.1991
{'key': 'e_1_2_6_30_1', 'first-page': '955', 'volume-title': 'Escherichia coli and Salmonella: Cellular and Molecular Biology', 'author': 'Wandersman C.', 'year': '1996'} / Escherichia coli and Salmonella: Cellular and Molecular Biology by Wandersman C. (1996)
10.1046/j.1365-2958.1998.00822.x
{'key': 'e_1_2_6_32_1', 'first-page': '3339', 'article-title': 'Genetic analysis of three additional fla genes in Salmonella typhimurium', 'volume': '130', 'author': 'Yamaguchi S.', 'year': '1984', 'journal-title': 'J Gen Microbiol'} / J Gen Microbiol / Genetic analysis of three additional fla genes in Salmonella typhimurium by Yamaguchi S. (1984)
10.1099/13500872-141-7-1715

Dates

Type	When
Created	22 years, 5 months ago (March 12, 2003, 6:29 a.m.)
Deposited	1 year, 9 months ago (Oct. 29, 2023, 11:58 a.m.)
Indexed	4 months ago (April 22, 2025, 1:43 p.m.)
Issued	25 years, 5 months ago (March 1, 2000)
Published	25 years, 5 months ago (March 1, 2000)
Published Online	23 years, 7 months ago (Jan. 18, 2002)
Published Print	25 years, 5 months ago (March 1, 2000)

Funders 0

None

BibTeX

@article{Minamino_2000, title={Interactions among components of the Salmonella flagellar export apparatus and its substrates}, volume={35}, ISSN={1365-2958}, url={http://dx.doi.org/10.1046/j.1365-2958.2000.01771.x}, DOI={10.1046/j.1365-2958.2000.01771.x}, number={5}, journal={Molecular Microbiology}, publisher={Wiley}, author={Minamino, Tohru and Macnab, Robert M.}, year={2000}, month=mar, pages={1052–1064} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        4,
        22
      ]
    ],
    "date-time": "2025-04-22T17:43:54Z",
    "timestamp": 1745343834481
  },
  "reference-count": 32,
  "publisher": "Wiley",
  "issue": "5",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2002,
            1,
            18
          ]
        ],
        "date-time": "2002-01-18T00:00:00Z",
        "timestamp": 1011312000000
      },
      "content-version": "vor",
      "delay-in-days": 688,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        2000,
        3
      ]
    ]
  },
  "abstract": "<jats:p>We have examined the cytoplasmic components (FliH, FliI and FliJ) of the type III flagellar protein export apparatus, plus the cytoplasmic domains (FlhA<jats:sub>C</jats:sub> and FlhB<jats:sub>C</jats:sub>) of two of its six membrane components. FliH, FlhA<jats:sub>C</jats:sub> and FliJ, when overproduced, caused inhibition of motility of wild\u2010type cells and inhibition of the export of substrates such as the hook protein FlgE. Co\u2010overproduction of FliH and FliI substantially relieved the inhibition caused by FliH, suggesting that it is excess free FliH that is inhibitory and that FliH and FliI form a complex. We purified His\u2010FLAG\u2010tagged versions of: (i) export components FliH, FliI, FliJ, FlhA<jats:sub>C</jats:sub> and FlhB<jats:sub>C</jats:sub>; (ii) rod/hook\u2010type export substrates FlgB (rod protein), FlgE (hook protein), FlgD (hook capping protein) and FliE (basal body protein); and (iii) filament\u2010type export substrates FlgK and FlgL (hook\u2013filament junction proteins) and FliC (flagellin). We tested for protein\u2013protein interactions by affinity blotting. In many cases, a given protein interacted with more than one other component, indicating that there are likely to be multiple dynamic interactions or interactions that involve more than two components. Interactions of FlhB<jats:sub>C</jats:sub> with rod/hook\u2010type substrates were strong, whereas those with filament\u2010type substrates were very weak; this may reflect the role of FlhB in substrate specificity switching. We propose a model for the flagellar export apparatus in which FlhA and FlhB and the other four integral membrane proteins of the apparatus form a complex at the base of the flagellar motor. A soluble complex of at least three proteins (FliH, FliI and FliJ) bind the protein to be exported and then interact with the complex at the motor to deliver the protein, which is then exported in an ATP\u2010dependent process mediated by FliI.</jats:p>",
  "DOI": "10.1046/j.1365-2958.2000.01771.x",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2003,
        3,
        12
      ]
    ],
    "date-time": "2003-03-12T11:29:38Z",
    "timestamp": 1047468578000
  },
  "page": "1052-1064",
  "source": "Crossref",
  "is-referenced-by-count": 178,
  "title": "Interactions among components of the <i>Salmonella</i> flagellar export apparatus and its substrates",
  "prefix": "10.1111",
  "volume": "35",
  "author": [
    {
      "given": "Tohru",
      "family": "Minamino",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Robert M.",
      "family": "Macnab",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2002,
        1,
        18
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_6_2_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.50.31981"
    },
    {
      "key": "e_1_2_6_3_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.1997.6412010.x"
    },
    {
      "key": "e_1_2_6_4_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.1999.01372.x"
    },
    {
      "key": "e_1_2_6_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0022-2836(05)80266-9"
    },
    {
      "key": "e_1_2_6_6_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0022-2836(90)90365-S"
    },
    {
      "key": "e_1_2_6_7_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/nbt1088-1204"
    },
    {
      "key": "e_1_2_6_8_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.2005.04758.x"
    },
    {
      "key": "e_1_2_6_9_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.8235660"
    },
    {
      "key": "e_1_2_6_10_1",
      "doi-asserted-by": "crossref",
      "first-page": "15758",
      "DOI": "10.1016/S0021-9258(17)36323-8",
      "article-title": "The covalent structure of the phase\u20101 flagellar filament protein of Salmonella typhimurium and its comparison with other flagellins",
      "volume": "260",
      "author": "Joys T.M.",
      "year": "1985",
      "journal-title": "J Biol Chem"
    },
    {
      "key": "e_1_2_6_11_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1996.0038"
    },
    {
      "key": "e_1_2_6_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.176.12.3598-3605.1994"
    },
    {
      "key": "e_1_2_6_13_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.176.24.7625-7629.1994"
    },
    {
      "key": "e_1_2_6_14_1",
      "first-page": "123",
      "volume-title": "Escherichia coli and Salmonella: Cellular and Molecular Biology",
      "author": "Macnab R.M.",
      "year": "1996"
    },
    {
      "key": "e_1_2_6_15_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.181.5.1388-1394.1999"
    },
    {
      "key": "e_1_2_6_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.176.24.7630-7637.1994"
    },
    {
      "key": "e_1_2_6_17_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.1999.01597.x"
    },
    {
      "key": "e_1_2_6_18_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1271/bbb.63.1301"
    },
    {
      "key": "e_1_2_6_19_1",
      "doi-asserted-by": "crossref",
      "first-page": "2298",
      "DOI": "10.1128/jb.174.7.2298-2304.1992",
      "article-title": "Characterization of the fliE genes of Escherichia coli and Salmonella typhimurium and identification of the FliE protein as a component of the flagellar hook\u2010basal body complex",
      "volume": "174",
      "author": "M\u00fcller V.",
      "year": "1992",
      "journal-title": "J Bacteriol"
    },
    {
      "key": "e_1_2_6_20_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.1998.00998.x"
    },
    {
      "key": "e_1_2_6_21_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.181.18.5808-5813.1999"
    },
    {
      "key": "e_1_2_6_22_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.181.5.1555-1561.1999"
    },
    {
      "key": "e_1_2_6_23_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.1992.tb01771.x"
    },
    {
      "key": "e_1_2_6_24_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.176.8.2272-2281.1994"
    },
    {
      "key": "e_1_2_6_25_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1742-4658.2006.05480.x"
    },
    {
      "key": "e_1_2_6_26_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.181.4.1059-1071.1999"
    },
    {
      "key": "e_1_2_6_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.179.17.5355-5365.1997"
    },
    {
      "key": "e_1_2_6_28_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jsbi.1998.4048"
    },
    {
      "key": "e_1_2_6_29_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.173.11.3564-3572.1991"
    },
    {
      "key": "e_1_2_6_30_1",
      "first-page": "955",
      "volume-title": "Escherichia coli and Salmonella: Cellular and Molecular Biology",
      "author": "Wandersman C.",
      "year": "1996"
    },
    {
      "key": "e_1_2_6_31_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.1998.00822.x"
    },
    {
      "key": "e_1_2_6_32_1",
      "first-page": "3339",
      "article-title": "Genetic analysis of three additional fla genes in Salmonella typhimurium",
      "volume": "130",
      "author": "Yamaguchi S.",
      "year": "1984",
      "journal-title": "J Gen Microbiol"
    },
    {
      "key": "e_1_2_6_33_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/13500872-141-7-1715"
    }
  ],
  "container-title": "Molecular Microbiology",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1046%2Fj.1365-2958.2000.01771.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.1365-2958.2000.01771.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        29
      ]
    ],
    "date-time": "2023-10-29T15:58:07Z",
    "timestamp": 1698595087000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://onlinelibrary.wiley.com/doi/10.1046/j.1365-2958.2000.01771.x"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2000,
        3
      ]
    ]
  },
  "references-count": 32,
  "journal-issue": {
    "issue": "5",
    "published-print": {
      "date-parts": [
        [
          2000,
          3
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1046/j.1365-2958.2000.01771.x"
  ],
  "URL": "http://dx.doi.org/10.1046/j.1365-2958.2000.01771.x",
  "relation": {},
  "ISSN": [
    "0950-382X",
    "1365-2958"
  ],
  "subject": [],
  "container-title-short": "Molecular Microbiology",
  "published": {
    "date-parts": [
      [
        2000,
        3
      ]
    ]
  }
}