DOI: 10.1073/pnas.98.4.1507. The quorum-sensing transcriptional regulator TraR requires its cognate signaling ligand for protein folding, protease resistance, and dimerization

The quorum-sensing transcriptional regulator TraR requires its cognate signaling ligand for protein folding, protease resistance, and dimerization

10.1073/pnas.98.4.1507

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Complexes between the quorum-sensing regulator TraR and its inducing ligand autoinducer (AAI) are soluble in Escherichia coli , whereas apo-TraR is almost completely insoluble. Here we show that the lack of soluble TraR is due in large part to rapid proteolysis, inasmuch as apo-TraR accumulated to high levels in an E. coli strain deficient in Clp and Lon proteases. In pulse labeling experiments, AAI protected TraR against proteolysis only when it was added before the radiolabel. This observation indicates that TraR proteins can productively bind AAI only during their own synthesis on polysomes, whereas fully synthesized apo-TraR proteins are not functional AAI receptors. Purified apo-TraR was rapidly degraded by trypsin to oligopeptides, whereas TraR–AAI complexes were more resistant to trypsin and were cleaved at discrete interdomain linkers, indicating that TraR requires AAI to attain its mature tertiary structure. TraR–AAI complexes eluted from a gel filtration column as dimers and bound DNA as dimers. In contrast, apo-TraR was monomeric, and incubation with AAI under a variety of conditions did not cause dimerization. We conclude that AAI is critical for the folding of nascent TraR protein into its mature tertiary structure and that full-length apo-TraR cannot productively bind AAI and is consequently targeted for rapid proteolysis.

Bibliography

Zhu, J., & Winans, S. C. (2001). The quorum-sensing transcriptional regulator TraR requires its cognate signaling ligand for protein folding, protease resistance, and dimerization. Proceedings of the National Academy of Sciences, 98(4), 1507â1512.

Authors 2

Jun Zhu (first)
Stephen C. Winans (additional)

References 29 Referenced 295

R Schleif Escherichia coli and Salmonella: Cellular and Molecular Biology, eds F C Neidhardt, R Curtiss, J L Ingraham, E C C Lin, K B Low, B Magasanik, W S Reznikoff, M Riley, M Schaechter, H E Umbarger (Am. Soc. Microbiol., Washington, DC), pp. 1300–1309 (1996). / Escherichia coli and Salmonella: Cellular and Molecular Biology by Schleif R (1996)
H Choy, S Adhya in Escherichia coli and Salmonella: Cellular and Molecular Biology, eds F C Neidhardt, R Curtiss, J L Ingraham, E C C Lin, K B Low, B Magasanik, W S Reznikoff, M Riley, M Schaechter, H E Umbarger (Am. Soc. Microbiol., Washington, DC), pp. 1287–1299 (1996). / in Escherichia coli and Salmonella: Cellular and Molecular Biology by Choy H (1996)
10.1128/9781555818319
10.1007/BF00039386
, eds G M Dunny, S C Winans (Am. Soc. Microbiol., Washington, DC Cell–Cell Signaling in Bacteria, 1999). / Cell–Cell Signaling in Bacteria by Dunny G M (1999)
10.1016/S1369-5274(98)80009-X
10.1023/A:1001178702503
10.1128/JB.182.14.3885-3895.2000
S C Winans, J Zhu, M I Moré Cell–Cell Signaling in Bacteria, eds G M Dunny, S C Winans (Am. Soc. Microbiol., Washington, DC), pp. 117–128 (1999). / Cell–Cell Signaling in Bacteria by Winans S C (1999)
10.1046/j.1365-2958.1998.01022.x
10.1073/pnas.91.26.12619
10.1073/pnas.96.9.4832
10.1093/emboj/19.4.631
10.1073/pnas.96.16.9009
10.1046/j.1365-2958.1998.00672.x
10.1128/jb.176.10.2796-2806.1994
10.1101/gad.12.9.1338
10.1101/gad.12.9.1348
10.1128/JB.181.12.3674-3680.1999
10.1128/JB.180.20.5398-5405.1998
10.1073/pnas.82.4.1074
10.1016/0378-1119(95)00584-1
10.1093/emboj/19.19.5212
10.1002/j.1460-2075.1987.tb02573.x
10.1073/pnas.96.11.6439
10.1146/annurev.genet.30.1.465
10.1073/pnas.77.6.3225
J W Roberts, C W Roberts Proc Natl Acad Sci USA 72, 4714–4751 (1975). / Proc Natl Acad Sci USA by Roberts J W (1975)
10.1126/science.286.5446.1888

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:45 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 5:37 p.m.)
Indexed	1 month, 1 week ago (July 19, 2025, 11:36 p.m.)
Issued	24 years, 6 months ago (Feb. 13, 2001)
Published	24 years, 6 months ago (Feb. 13, 2001)
Published Online	24 years, 6 months ago (Feb. 13, 2001)
Published Print	24 years, 6 months ago (Feb. 13, 2001)

Funders 0

None

BibTeX

@article{Zhu_2001, title={The quorum-sensing transcriptional regulator TraR requires its cognate signaling ligand for protein folding, protease resistance, and dimerization}, volume={98}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.98.4.1507}, DOI={10.1073/pnas.98.4.1507}, number={4}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Zhu, Jun and Winans, Stephen C.}, year={2001}, month=feb, pages={1507–1512} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        20
      ]
    ],
    "date-time": "2025-07-20T03:36:04Z",
    "timestamp": 1752982564541
  },
  "reference-count": 29,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "4",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2001,
        2,
        13
      ]
    ]
  },
  "abstract": "<jats:p>\n            Complexes between the quorum-sensing regulator TraR and its\n inducing ligand autoinducer (AAI) are soluble in\n            <jats:italic>Escherichia\n coli</jats:italic>\n            , whereas apo-TraR is almost completely insoluble. Here we\n show that the lack of soluble TraR is due in large part to rapid\n proteolysis, inasmuch as apo-TraR accumulated to high levels in an\n            <jats:italic>E. coli</jats:italic>\n            strain deficient in Clp and Lon proteases. In\n pulse labeling experiments, AAI protected TraR against proteolysis only\n when it was added before the radiolabel. This observation indicates\n that TraR proteins can productively bind AAI only during their own\n synthesis on polysomes, whereas fully synthesized apo-TraR proteins are\n not functional AAI receptors. Purified apo-TraR was rapidly degraded by\n trypsin to oligopeptides, whereas TraR\u2013AAI complexes were more\n resistant to trypsin and were cleaved at discrete interdomain linkers,\n indicating that TraR requires AAI to attain its mature tertiary\n structure. TraR\u2013AAI complexes eluted from a gel filtration column as\n dimers and bound DNA as dimers. In contrast, apo-TraR was monomeric,\n and incubation with AAI under a variety of conditions did not cause\n dimerization. We conclude that AAI is critical for the folding of\n nascent TraR protein into its mature tertiary structure and that\n full-length apo-TraR cannot productively bind AAI and is consequently\n targeted for rapid proteolysis.\n          </jats:p>",
  "DOI": "10.1073/pnas.98.4.1507",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T14:45:09Z",
    "timestamp": 1027694709000
  },
  "page": "1507-1512",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 295,
  "title": "The quorum-sensing transcriptional regulator TraR requires its cognate signaling ligand for protein folding, protease resistance, and dimerization",
  "prefix": "10.1073",
  "volume": "98",
  "author": [
    {
      "given": "Jun",
      "family": "Zhu",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Department of Microbiology, Cornell University, Ithaca, NY 14853"
        }
      ]
    },
    {
      "given": "Stephen C.",
      "family": "Winans",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Microbiology, Cornell University, Ithaca, NY 14853"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        2001,
        2,
        13
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_3_1_2",
      "first-page": "1300",
      "volume-title": "Escherichia coli and Salmonella: Cellular and Molecular Biology",
      "author": "Schleif R",
      "year": "1996",
      "unstructured": "R Schleif Escherichia coli and Salmonella: Cellular and Molecular Biology, eds F C Neidhardt, R Curtiss, J L Ingraham, E C C Lin, K B Low, B Magasanik, W S Reznikoff, M Riley, M Schaechter, H E Umbarger (Am. Soc. Microbiol., Washington, DC), pp. 1300\u20131309 (1996)."
    },
    {
      "key": "e_1_3_3_2_2",
      "first-page": "1287",
      "volume-title": "in Escherichia coli and Salmonella: Cellular and Molecular Biology",
      "author": "Choy H",
      "year": "1996",
      "unstructured": "H Choy, S Adhya in Escherichia coli and Salmonella: Cellular and Molecular Biology, eds F C Neidhardt, R Curtiss, J L Ingraham, E C C Lin, K B Low, B Magasanik, W S Reznikoff, M Riley, M Schaechter, H E Umbarger (Am. Soc. Microbiol., Washington, DC), pp. 1287\u20131299 (1996)."
    },
    {
      "key": "e_1_3_3_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/9781555818319"
    },
    {
      "key": "e_1_3_3_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00039386"
    },
    {
      "key": "e_1_3_3_5_2",
      "volume-title": "Cell\u2013Cell Signaling in Bacteria",
      "author": "Dunny G M",
      "year": "1999",
      "unstructured": ", eds G M Dunny, S C Winans (Am. Soc. Microbiol., Washington, DC Cell\u2013Cell Signaling in Bacteria, 1999)."
    },
    {
      "key": "e_1_3_3_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1369-5274(98)80009-X"
    },
    {
      "key": "e_1_3_3_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1023/A:1001178702503"
    },
    {
      "key": "e_1_3_3_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.182.14.3885-3895.2000"
    },
    {
      "key": "e_1_3_3_9_2",
      "first-page": "117",
      "volume-title": "Cell\u2013Cell Signaling in Bacteria",
      "author": "Winans S C",
      "year": "1999",
      "unstructured": "S C Winans, J Zhu, M I Mor\u00e9 Cell\u2013Cell Signaling in Bacteria, eds G M Dunny, S C Winans (Am. Soc. Microbiol., Washington, DC), pp. 117\u2013128 (1999)."
    },
    {
      "key": "e_1_3_3_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.1998.01022.x"
    },
    {
      "key": "e_1_3_3_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.91.26.12619"
    },
    {
      "key": "e_1_3_3_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.96.9.4832"
    },
    {
      "key": "e_1_3_3_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/19.4.631"
    },
    {
      "key": "e_1_3_3_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.96.16.9009"
    },
    {
      "key": "e_1_3_3_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.1998.00672.x"
    },
    {
      "key": "e_1_3_3_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.176.10.2796-2806.1994"
    },
    {
      "key": "e_1_3_3_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.12.9.1338"
    },
    {
      "key": "e_1_3_3_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.12.9.1348"
    },
    {
      "key": "e_1_3_3_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.181.12.3674-3680.1999"
    },
    {
      "key": "e_1_3_3_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JB.180.20.5398-5405.1998"
    },
    {
      "key": "e_1_3_3_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.82.4.1074"
    },
    {
      "key": "e_1_3_3_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0378-1119(95)00584-1"
    },
    {
      "key": "e_1_3_3_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/19.19.5212"
    },
    {
      "key": "e_1_3_3_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1987.tb02573.x"
    },
    {
      "key": "e_1_3_3_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.96.11.6439"
    },
    {
      "key": "e_1_3_3_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.genet.30.1.465"
    },
    {
      "key": "e_1_3_3_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.77.6.3225"
    },
    {
      "key": "e_1_3_3_28_2",
      "first-page": "4714",
      "volume": "72",
      "author": "Roberts J W",
      "year": "1975",
      "unstructured": "J W Roberts, C W Roberts Proc Natl Acad Sci USA 72, 4714\u20134751 (1975).",
      "journal-title": "Proc Natl Acad Sci USA"
    },
    {
      "key": "e_1_3_3_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.286.5446.1888"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.98.4.1507",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        4,
        13
      ]
    ],
    "date-time": "2022-04-13T21:37:36Z",
    "timestamp": 1649885856000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.98.4.1507"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2001,
        2,
        13
      ]
    ]
  },
  "references-count": 29,
  "journal-issue": {
    "issue": "4",
    "published-print": {
      "date-parts": [
        [
          2001,
          2,
          13
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.98.4.1507"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.98.4.1507",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        2001,
        2,
        13
      ]
    ]
  },
  "assertion": [
    {
      "value": "2000-09-25",
      "order": 0,
      "name": "received",
      "label": "Received",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "2001-02-13",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}