DOI: 10.1046/j.1365-2958.2003.03481.x. Medicinal genetics approach towards identifying the molecular target of a novel inhibitor of fungal cell wall assembly

Medicinal genetics approach towards identifying the molecular target of a novel inhibitor of fungal cell wall assembly

10.1046/j.1365-2958.2003.03481.x

Crossref journal-article

Wiley

Molecular Microbiology (311)

Abstract

SummaryGlycosylphosphatidylinositol (GPI)‐anchored cell wall mannoproteins are required for the adhesion of pathogenic fungi, such as Candida albicans, to human epithelium. Small molecular inhibitors of the cell surface presentation of GPI‐anchored mannoproteins would be promising candidate drugs to block the establishment of fungal infections. Here, we describe a medicinal genetics approach to identifying the gene encoding a novel target protein that is required for the localization of GPI‐anchored cell wall mannoproteins. By means of a yeast cell‐based screening procedure, we discovered a compound, 1‐[4‐butylbenzyl]isoquinoline (BIQ), that inhibits cell wall localization of GPI‐anchored mannoproteins in Saccharomyces cerevisiae. Treatment of C. albicans cells with this compound resulted in reduced adherence to a rat intestine epithelial cell monolayer. A previously uncharacterized gene YJL091c, named GWT1, was cloned as a dosage‐dependent suppressor of the BIQ‐induced phenotypes. GWT1 knock‐out cells showed similar phenotypes to BIQ‐treated wild‐type cells in terms of cell wall structure and transcriptional profiles. Two different mutants resistant to BIQ each contained a single missense mutation in the coding region of the GWT1 gene. These results all suggest that the GWT1 gene product is the primary target of the compound.

Bibliography

Tsukahara, K., Hata, K., Nakamoto, K., Sagane, K., Watanabe, N., Kuromitsu, J., Kai, J., Tsuchiya, M., Ohba, F., Jigami, Y., Yoshimatsu, K., & Nagasu, T. (2003). Medicinal genetics approach towards identifying the molecular target of a novel inhibitor of fungal cell wall assembly. Molecular Microbiology, 48(4), 1029â1042. Portico.

Authors 12

Kappei Tsukahara (first)
Katsura Hata (additional)
Kazutaka Nakamoto (additional)
Koji Sagane (additional)
Nao‐aki Watanabe (additional)
Junro Kuromitsu (additional)
Junko Kai (additional)
Mamiko Tsuchiya (additional)
Fuminori Ohba (additional)
Yoshifumi Jigami (additional)
Kentaro Yoshimatsu (additional)
Takeshi Nagasu (additional)

References 38 Referenced 100

10.1126/science.8284673
10.1091/mbc.10.4.1043
10.1073/pnas.261574398
10.1111/j.1574-6976.2002.tb00613.x
10.1016/S0968-0004(99)01425-5
10.1038/85677
10.1046/j.1365-2958.2002.02832.x
10.1016/S1359-6446(99)01430-0
{'key': 'e_1_2_6_10_1', 'first-page': '515', 'article-title': 'Fungal infection rates are increasing', 'volume': '10', 'author': 'Fox J.L.', 'year': '1993', 'journal-title': 'Am Soc Microbiol News'} / Am Soc Microbiol News / Fungal infection rates are increasing by Fox J.L. (1993)
10.1128/IAI.66.4.1783-1786.1998 / Infect Immun / Expression of the Candida albicans gene ALS1. Saccharomyces cerevisiae induces adherence to endothelial and epithelial cells by Fu Y. (1998)
10.1128/IAI.65.12.5289-5294.1997
10.1128/AAC.36.10.2239
10.1038/6791
{'key': 'e_1_2_6_15_1', 'first-page': '12', 'volume-title': 'Guide to Yeast Genetics and Molecular Biology.', 'author': 'Guthrie C.', 'year': '1991'} / Guide to Yeast Genetics and Molecular Biology. by Guthrie C. (1991)
10.1128/AAC.40.10.2243
10.1203/00006450-199604000-00001
10.1111/j.1365-2958.1995.tb02219.x
10.1271/bbb.57.1686
10.1111/j.1432-1033.1995.0845d.x
10.1093/glycob/6.3.337
10.1016/S1367-5931(00)00151-4
10.1074/jbc.272.28.17762
10.1002/(SICI)1097-0061(199807)14:10<935::AID-YEA289>3.0.CO;2-9
10.1038/3282
10.1128/MCB.15.10.5671
10.1002/yea.320110709
10.1126/science.285.5427.591
10.1086/344058
10.1128/JCM.40.10.3551-3557.2002
10.1111/j.1574-6968.1998.tb13006.x
10.1073/pnas.80.22.6750
10.1093/nar/18.10.3091
10.1002/yea.320090410
10.1093/genetics/122.1.19
10.1126/science.283.5407.1535
10.1016/S0378-1097(96)00440-5
10.1002/yea.320101310
10.1038/nbt1297-1359

Dates

Type	When
Created	22 years, 3 months ago (May 8, 2003, 10:51 a.m.)
Deposited	1 year, 10 months ago (Oct. 13, 2023, 8:58 p.m.)
Indexed	1 week, 6 days ago (Aug. 21, 2025, 1:43 p.m.)
Issued	22 years, 4 months ago (May 1, 2003)
Published	22 years, 4 months ago (May 1, 2003)
Published Online	22 years, 3 months ago (May 8, 2003)
Published Print	22 years, 4 months ago (May 1, 2003)

Funders 0

None

BibTeX

@article{Tsukahara_2003, title={Medicinal genetics approach towards identifying the molecular target of a novel inhibitor of fungal cell wall assembly}, volume={48}, ISSN={1365-2958}, url={http://dx.doi.org/10.1046/j.1365-2958.2003.03481.x}, DOI={10.1046/j.1365-2958.2003.03481.x}, number={4}, journal={Molecular Microbiology}, publisher={Wiley}, author={Tsukahara, Kappei and Hata, Katsura and Nakamoto, Kazutaka and Sagane, Koji and Watanabe, Nao‐aki and Kuromitsu, Junro and Kai, Junko and Tsuchiya, Mamiko and Ohba, Fuminori and Jigami, Yoshifumi and Yoshimatsu, Kentaro and Nagasu, Takeshi}, year={2003}, month=may, pages={1029–1042} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        21
      ]
    ],
    "date-time": "2025-08-21T17:43:55Z",
    "timestamp": 1755798235488
  },
  "reference-count": 38,
  "publisher": "Wiley",
  "issue": "4",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2003,
            5,
            8
          ]
        ],
        "date-time": "2003-05-08T00:00:00Z",
        "timestamp": 1052352000000
      },
      "content-version": "vor",
      "delay-in-days": 7,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        2003,
        5
      ]
    ]
  },
  "abstract": "<jats:title>Summary</jats:title><jats:p>Glycosylphosphatidylinositol (GPI)\u2010anchored cell wall mannoproteins are required for the adhesion of pathogenic fungi, such as <jats:italic>Candida albicans</jats:italic>, to human epithelium. Small molecular inhibitors of the cell surface presentation of GPI\u2010anchored mannoproteins would be promising candidate drugs to block the establishment of fungal infections. Here, we describe a medicinal genetics approach to identifying the gene encoding a novel target protein that is required for the localization of GPI\u2010anchored cell wall mannoproteins. By means of a yeast cell\u2010based screening procedure, we discovered a compound, 1\u2010[4\u2010butylbenzyl]isoquinoline (BIQ), that inhibits cell wall localization of GPI\u2010anchored mannoproteins in <jats:italic>Saccharomyces cerevisiae</jats:italic>. Treatment of <jats:italic>C. albicans</jats:italic> cells with this compound resulted in reduced adherence to a rat intestine epithelial cell monolayer. A previously uncharacterized gene <jats:italic>YJL091c</jats:italic>, named <jats:italic>GWT1</jats:italic>, was cloned as a dosage\u2010dependent suppressor of the BIQ\u2010induced phenotypes. <jats:italic>GWT1</jats:italic> knock\u2010out cells showed similar phenotypes to BIQ\u2010treated wild\u2010type cells in terms of cell wall structure and transcriptional profiles. Two different mutants resistant to BIQ each contained a single missense mutation in the coding region of the <jats:italic>GWT1</jats:italic> gene. These results all suggest that the <jats:italic>GWT1</jats:italic> gene product is the primary target of the compound.</jats:p>",
  "DOI": "10.1046/j.1365-2958.2003.03481.x",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2003,
        5,
        8
      ]
    ],
    "date-time": "2003-05-08T14:51:59Z",
    "timestamp": 1052405519000
  },
  "page": "1029-1042",
  "source": "Crossref",
  "is-referenced-by-count": 100,
  "title": "Medicinal genetics approach towards identifying the molecular target of a novel inhibitor of fungal cell wall assembly",
  "prefix": "10.1111",
  "volume": "48",
  "author": [
    {
      "given": "Kappei",
      "family": "Tsukahara",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Katsura",
      "family": "Hata",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Kazutaka",
      "family": "Nakamoto",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Koji",
      "family": "Sagane",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Nao\u2010aki",
      "family": "Watanabe",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Junro",
      "family": "Kuromitsu",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Junko",
      "family": "Kai",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Mamiko",
      "family": "Tsuchiya",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Fuminori",
      "family": "Ohba",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Yoshifumi",
      "family": "Jigami",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Kentaro",
      "family": "Yoshimatsu",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Takeshi",
      "family": "Nagasu",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2003,
        5,
        8
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_6_2_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.8284673"
    },
    {
      "key": "e_1_2_6_3_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.10.4.1043"
    },
    {
      "key": "e_1_2_6_4_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.261574398"
    },
    {
      "key": "e_1_2_6_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1574-6976.2002.tb00613.x"
    },
    {
      "key": "e_1_2_6_6_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0968-0004(99)01425-5"
    },
    {
      "key": "e_1_2_6_7_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/85677"
    },
    {
      "key": "e_1_2_6_8_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2958.2002.02832.x"
    },
    {
      "key": "e_1_2_6_9_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1359-6446(99)01430-0"
    },
    {
      "key": "e_1_2_6_10_1",
      "first-page": "515",
      "article-title": "Fungal infection rates are increasing",
      "volume": "10",
      "author": "Fox J.L.",
      "year": "1993",
      "journal-title": "Am Soc Microbiol News"
    },
    {
      "key": "e_1_2_6_11_1",
      "doi-asserted-by": "crossref",
      "first-page": "1783",
      "DOI": "10.1128/IAI.66.4.1783-1786.1998",
      "article-title": "Expression of the Candida albicans gene ALS1. Saccharomyces cerevisiae induces adherence to endothelial and epithelial cells",
      "volume": "66",
      "author": "Fu Y.",
      "year": "1998",
      "journal-title": "Infect Immun"
    },
    {
      "key": "e_1_2_6_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/IAI.65.12.5289-5294.1997"
    },
    {
      "key": "e_1_2_6_13_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/AAC.36.10.2239"
    },
    {
      "key": "e_1_2_6_14_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/6791"
    },
    {
      "key": "e_1_2_6_15_1",
      "first-page": "12",
      "volume-title": "Guide to Yeast Genetics and Molecular Biology.",
      "author": "Guthrie C.",
      "year": "1991"
    },
    {
      "key": "e_1_2_6_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/AAC.40.10.2243"
    },
    {
      "key": "e_1_2_6_17_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1203/00006450-199604000-00001"
    },
    {
      "key": "e_1_2_6_18_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1365-2958.1995.tb02219.x"
    },
    {
      "key": "e_1_2_6_19_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1271/bbb.57.1686"
    },
    {
      "key": "e_1_2_6_20_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1432-1033.1995.0845d.x"
    },
    {
      "key": "e_1_2_6_21_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/glycob/6.3.337"
    },
    {
      "key": "e_1_2_6_22_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1367-5931(00)00151-4"
    },
    {
      "key": "e_1_2_6_23_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.28.17762"
    },
    {
      "key": "e_1_2_6_24_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/(SICI)1097-0061(199807)14:10<935::AID-YEA289>3.0.CO;2-9"
    },
    {
      "key": "e_1_2_6_25_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/3282"
    },
    {
      "key": "e_1_2_6_26_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MCB.15.10.5671"
    },
    {
      "key": "e_1_2_6_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/yea.320110709"
    },
    {
      "key": "e_1_2_6_28_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.285.5427.591"
    },
    {
      "key": "e_1_2_6_29_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1086/344058"
    },
    {
      "key": "e_1_2_6_30_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/JCM.40.10.3551-3557.2002"
    },
    {
      "key": "e_1_2_6_31_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1574-6968.1998.tb13006.x"
    },
    {
      "key": "e_1_2_6_32_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.80.22.6750"
    },
    {
      "key": "e_1_2_6_33_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/18.10.3091"
    },
    {
      "key": "e_1_2_6_34_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/yea.320090410"
    },
    {
      "key": "e_1_2_6_35_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/genetics/122.1.19"
    },
    {
      "key": "e_1_2_6_36_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.283.5407.1535"
    },
    {
      "key": "e_1_2_6_37_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0378-1097(96)00440-5"
    },
    {
      "key": "e_1_2_6_38_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/yea.320101310"
    },
    {
      "key": "e_1_2_6_39_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/nbt1297-1359"
    }
  ],
  "container-title": "Molecular Microbiology",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1046%2Fj.1365-2958.2003.03481.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.1365-2958.2003.03481.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        14
      ]
    ],
    "date-time": "2023-10-14T00:58:43Z",
    "timestamp": 1697245123000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://onlinelibrary.wiley.com/doi/10.1046/j.1365-2958.2003.03481.x"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2003,
        5
      ]
    ]
  },
  "references-count": 38,
  "journal-issue": {
    "issue": "4",
    "published-print": {
      "date-parts": [
        [
          2003,
          5
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1046/j.1365-2958.2003.03481.x"
  ],
  "URL": "http://dx.doi.org/10.1046/j.1365-2958.2003.03481.x",
  "relation": {},
  "ISSN": [
    "0950-382X",
    "1365-2958"
  ],
  "subject": [],
  "container-title-short": "Molecular Microbiology",
  "published": {
    "date-parts": [
      [
        2003,
        5
      ]
    ]
  }
}