Inhibiting GPI Anchor Biosynthesis in Fungi Stresses the Endoplasmic Reticulum and Enhances Immunogenicity
Open accessAbstract
In fungi, the anchoring of proteins to the plasma membrane via their covalent attachment to glycosylphosphatidylinositol (GPI) is essential and thus provides a valuable point of attack for the development of antifungal therapeutics. Unfortunately, studying the underlying biology of GPI-anchor synthesis is difficult, especially in medically relevant fungal pathogens because they are not genetically tractable. Compounding difficulties, many of the genes in this pathway are essential in Saccharomyces cerevisiae. Here, we report the discovery of a new small molecule christened gepinacin (for GPI acylation inhibitor) which selectively inhibits Gwt1, a critical acyltransferase required for the biosynthesis of fungal GPI anchors. After delineating the target specificity of gepinacin using genetic and biochemical techniques, we used it to probe key, therapeutically relevant consequences of disrupting GPI anchor metabolism in fungi. We found that, unlike all three major classes of antifungals in current use, the direct antimicrobial activity of this compound results predominantly from its ability to induce overwhelming stress to the endoplasmic reticulum. Gepinacin did not affect the viability of mammalian cells nor did it inhibit their orthologous acyltransferase. This enabled its use in co-culture experiments to examine Gwt1's effects on host-pathogen interactions. In isolates of Candida albicans, the most common fungal pathogen in humans, exposure to gepinacin at sublethal concentrations impaired filamentation and unmasked cell wall β-glucan to stimulate a pro-inflammatory cytokine response in macrophages. Gwt1 is a promising antifungal drug target, and gepanacin is a useful probe for studying how disrupting GPI-anchor synthesis impairs viability and alters host-pathogen interactions in genetically intractable fungi.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Inhibition of Gwt1 results in pleiotropic effects on the fungal cell including alterations in fungal adherence to surfaces, inhibition of biofilm and germ tube formation, and subsequently results in severe growth defects and yeast lethality.
“Inhibition of Gwt1 results in pleiotropic effects on the fungal cell including alterations in fungal adherence to surfaces, inhibition of biofilm and germ tube formation, and subsequently results in severe growth defects and yeast lethality (15).”
From Clinical Efficacy and Safety of a Novel Antifungal, Fosmanogepix, in Patients with Candidemia Caused by Candida auris <i>:</i> Results from a Phase 2 Trial (2023) · cites this paper as [15]
Cites
The Unfolded Protein Response: From Stress Pathway to Homeostatic Regulation
Walter, P., & Ron, D. (2011) · Science · doi:10.1126/science.1209038 · Cited by 5,893 · see citing works
Functional profiling of the Saccharomyces cerevisiae genome
Giaever, G., Chu, A. M., Ni, L., Connelly, C., Riles, L., Véronneau, S., Dow, S., Lucau-Danila, A., Anderson, K., André, B., et al. (2002) · Nature · doi:10.1038/nature00935 · Cited by 4,316 · see citing works
Global analysis of protein localization in budding yeast
Huh, W.-K., Falvo, J. V., Gerke, L. C., Carroll, A. S., Howson, R. W., Weissman, J. S., & O'Shea, E. K. (2003) · Nature · doi:10.1038/nature02026 · Cited by 4,248 · see citing works
Epidemiology of Invasive Candidiasis: a Persistent Public Health Problem
Pfaller, M. A., & Diekema, D. J. (2007) · Clinical Microbiology Reviews · doi:10.1128/cmr.00029-06 · Cited by 4,174 · see citing works
Cox, J. S., Shamu, C. E., & Walter, P. (1993) · Cell · doi:10.1016/0092-8674(93)90648-a · Cited by 1,240 · see citing works
Growth of Candida albicans hyphae
Sudbery, P. E. (2011) · Nature Reviews Microbiology · doi:10.1038/nrmicro2636 · Cited by 1,110 · see citing works
Molecular Evidence for the Early Colonization of Land by Fungi and Plants
Heckman, D. S., Geiser, D. M., Eidell, B. R., Stauffer, R. L., Kardos, N. L., & Hedges, S. B. (2001) · Science · doi:10.1126/science.1061457 · Cited by 1,020 · see citing works
Suppression of Hyphal Formation in Candida albicans by Mutation of a STE12 Homolog
Liu, H., Köhler, J. R., & Fink, G. R. (1994) · Science · doi:10.1126/science.7992058 · Cited by 788 · see citing works
Noble, S. M., French, S., Kohn, L. A., Chen, V., & Johnson, A. D. (2010) · Nature Genetics · doi:10.1038/ng.605 · Cited by 678 · see citing works
Regulatory circuitry governing fungal development, drug resistance, and disease.
Shapiro, R. S., Robbins, N., & Cowen, L. E. (2011) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.00045-10 · Cited by 576 · see citing works
A suite of Gateway® cloning vectors for high‐throughput genetic analysis in Saccharomyces cerevisiae
Alberti, S., Gitler, A. D., & Lindquist, S. L. (2007) · Yeast · doi:10.1002/yea.1502 · Cited by 518 · see citing works
Self-citationHeifetz, A. H., Keenan, R. W., & Elbein, A. D. (1979) · Biochemistry · doi:10.1021/bi00578a008 · Cited by 446 · see citing works
Pollard, M. G., Travers, K. J., & Weissman, J. S. (1998) · Molecular Cell · doi:10.1016/s1097-2765(00)80018-0 · Cited by 441 · see citing works
An insight into the antifungal pipeline: selected new molecules and beyond
Ostrosky-Zeichner, L., Casadevall, A., Galgiani, J. N., Odds, F. C., & Rex, J. H. (2010) · Nature Reviews Drug Discovery · doi:10.1038/nrd3074 · Cited by 432 · see citing works
Orlean, P., & Menon, A. K. (2007) · Journal of Lipid Research · doi:10.1194/jlr.r700002-jlr200 · Cited by 403 · see citing works
The Unfolding Tale of the Unfolded Protein Response
Ma, Y., & Hendershot, L. M. (2001) · Cell · doi:10.1016/s0092-8674(01)00623-7 · Cited by 401 · see citing works
A Drug-Sensitive Genetic Network Masks Fungi from the Immune System
Wheeler, R. T., & Fink, G. R. (2006) · PLoS Pathogens · doi:10.1371/journal.ppat.0020035 · Cited by 360 · see citing works
Dynamic, Morphotype-Specific Candida albicans β-Glucan Exposure during Infection and Drug Treatment
Wheeler, R. T., Kombe, D., Agarwala, S. D., & Fink, G. R. (2008) · PLoS Pathogens · doi:10.1371/journal.ppat.1000227 · Cited by 309 · see citing works
Vacuole Biogenesis in Saccharomyces cerevisiae : Protein Transport Pathways to the Yeast Vacuole
Bryant, N. J., & Stevens, T. H. (1998) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.62.1.230-247.1998 · Cited by 283 · see citing works
An integrated platform of genomic assays reveals small-molecule bioactivities
Hoon, S. S., Smith, A. M., Wallace, I. M., Suresh, S., Miranda, M., Fung, E., Proctor, M. J., Shokat, K. M., Zhang, C., Davis, R. W., et al. (2008) · Nature Chemical Biology · doi:10.1038/nchembio.100 · Cited by 201 · see citing works
<i>In Vitro</i> Activity of E1210, a Novel Antifungal, against Clinically Important Yeasts and Molds
Miyazaki, M., Horii, T., Hata, K., Watanabe, N.-A., Nakamoto, K., Tanaka, K., Shirotori, S., Murai, N., Inoue, S., Matsukura, M., et al. (2011) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.00291-11 · Cited by 200 · see citing works
The p24 family and selective transport processes at the ER—Golgi interface
Strating, J. R. P. M., & Martens, G. J. M. (2009) · Biology of the Cell · doi:10.1042/bc20080233 · Cited by 192 · see citing works
Watanabe, N.-A., Miyazaki, M., Horii, T., Sagane, K., Tsukahara, K., & Hata, K. (2012) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.00731-11 · Cited by 169 · see citing works
Klis, F. M., Sosinska, G. J., de Groot, P. W. J., & Brul, S. (2009) · FEMS Yeast Research · doi:10.1111/j.1567-1364.2009.00541.x · Cited by 165 · see citing works
Candida albicans cell wall glycans, host receptors and responses: elements for a decisive crosstalk
Poulain, D., & Jouault, T. (2004) · Current Opinion in Microbiology · doi:10.1016/j.mib.2004.06.011 · Cited by 153 · see citing works
Seipin: from human disease to molecular mechanism
Cartwright, B. R., & Goodman, J. M. (2012) · Journal of Lipid Research · doi:10.1194/jlr.r023754 · Cited by 152 · see citing works
Outwitting Multidrug Resistance to Antifungals
Monk, B. C., & Goffeau, A. (2008) · Science · doi:10.1126/science.1159746 · Cited by 151 · see citing works
Evolution of Drug Resistance in <i>Candida Albicans</i>
Cowen, L. E., Anderson, J. B., & Kohn, L. M. (2002) · Annual Review of Microbiology · doi:10.1146/annurev.micro.56.012302.160907 · Cited by 151 · see citing works
Hata, K., Horii, T., Miyazaki, M., Watanabe, N.-A., Okubo, M., Sonoda, J., Nakamoto, K., Tanaka, K., Shirotori, S., Murai, N., et al. (2011) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.00366-11 · Cited by 149 · see citing works
Castillon, G. A., Aguilera-Romero, A., Manzano‐Lopez, J., et al. (2011) · Molecular Biology of the Cell · doi:10.1091/mbc.e11-04-0294 · Cited by 133 · see citing works
GWT1 Gene Is Required for Inositol Acylation of Glycosylphosphatidylinositol Anchors in Yeast
Umemura, M., Okamoto, M., Nakayama, K.-I., Sagane, K., Tsukahara, K., Hata, K., & Jigami, Y. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m301044200 · Cited by 127 · see citing works
Tsukahara, K., Hata, K., Nakamoto, K., Sagane, K., Watanabe, N.-A., Kuromitsu, J., Kai, J.-K., Tsuchiya, M., Ohba, F., Jigami, Y., et al. (2003) · Molecular Microbiology · doi:10.1046/j.1365-2958.2003.03481.x · Cited by 117 · see citing works
A unique and universal molecular barcode array
Pierce, S. E., Fung, E., Jaramillo, D. F., Chu, A. M., Davis, R. W., Nislow, C., & Giaever, G. N. (2006) · Nature Methods · doi:10.1038/nmeth905 · Cited by 115 · see citing works
PIG-W Is Critical for Inositol Acylation but Not for Flipping of Glycosylphosphatidylinositol-Anchor
Murakami, Y., Siripanyapinyo, U., Hong, Y., Kang, J. Y., Ishihara, S., Nakakuma, H., Maeda, Y., & Kinoshita, T. (2003) · Molecular Biology of the Cell · doi:10.1091/mbc.e03-03-0193 · Cited by 110 · see citing works
Hu, Y., Rolfs, A., Bhullar, B., Murthy, T. V. S., Zhu, C., Berger, M. F., Camargo, A. A., Kelley, F., McCarron, S., Jepson, D., et al. (2007) · Genome Research · doi:10.1101/gr.6037607 · Cited by 109 · see citing works
White, T., Pfaller, M., Rinaldi, M., Smith, J., & Redding, S. (1997) · Oral Diseases · doi:10.1111/j.1601-0825.1997.tb00336.x · Cited by 107 · see citing works
Fujita, M., Yoko‐o, T., & Jigami, Y. (2005) · Molecular Biology of the Cell · doi:10.1091/mbc.e05-05-0443 · Cited by 95 · see citing works
Shen, J., Cowen, L. E., Griffin, A. M., Chan, L. Y., & Köhler, J. R. (2008) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0809147105 · Cited by 70 · see citing works
GPI7 affects cell-wall protein anchorage in Saccharomyces cerevisiae and Candida albicans
Richard, M. L., de Groot, P. W. J., Courtin, O., Poulain, D., Klis, F. M., & Gaillardin, C. M. (2002) · Microbiology · doi:10.1099/00221287-148-7-2125 · Cited by 58 · see citing works
Synthesis and evaluation of novel antifungal agents-quinoline and pyridine amide derivatives
Nakamoto, K., TSUKADA, I., Tanaka, K., Matsukura, M., Haneda, T., Inoue, S., Murai, N., Abe, S., Ueda, N., Miyazaki, M., et al. (2010) · Bioorganic & Medicinal Chemistry Letters · doi:10.1016/j.bmcl.2010.06.005 · Cited by 46 · see citing works
Fankhauser, C., & Conzelmann, A. (1991) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1991.tb15723.x · Cited by 41 · see citing works
Sagane, K., Umemura, M., Ogawa‐Mitsuhashi, K., et al. (2011) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.193490 · Cited by 37 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
The antifungal pipeline: a reality check
Perfect, J. R. (2017) · Nature Reviews Drug Discovery · doi:10.1038/nrd.2017.46 · Cited by 866 · see citing works
Antifungal Drug Development: Challenges, Unmet Clinical Needs, and New Approaches
Roemer, T., & Krysan, D. J. (2014) · Cold Spring Harbor Perspectives in Medicine · doi:10.1101/cshperspect.a019703 · Cited by 593 · see citing works
Antifungal Drugs: The Current Armamentarium and Development of New Agents
Robbins, N., Wright, G. D., & Cowen, L. E. (2016) · Microbiology Spectrum · doi:10.1128/microbiolspec.funk-0002-2016 · Cited by 271 · see citing works
Fungal Cell Wall: Emerging Antifungals and Drug Resistance.
Lima, S. L., Colombo, A. L., & de Almeida Junior, J. N. (2019) · Frontiers in Microbiology · doi:10.3389/fmicb.2019.02573 · Cited by 223 · see citing works
Shaw, K. J., & Ibrahim, A. S. (2020) · Journal of Fungi · doi:10.3390/jof6040239 · Cited by 229 · see citing works
Hager, C. L., Larkin, E. L., Long, L., Zohra Abidi, F., Shaw, K. J., & Ghannoum, M. A. (2018) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.02319-17 · Cited by 139 · see citing works
Shaw, K. J., Schell, W. A., Covel, J., Duboc, G., Giamberardino, C., Kapoor, M., Moloney, M., Soltow, Q. A., Tenor, J. L., Toffaletti, D. L., et al. (2018) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.00523-18 · Cited by 121 · see citing works
Vazquez, J. A., Pappas, P. G., Boffard, K., Paruk, F., Bien, P. A., Tawadrous, M., Ople, E., Wedel, P., Oborska, I., & Hodges, M. R. (2023) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.01419-22 · Cited by 92 · see citing works
Exploiting mitochondria as targets for the development of new antifungals
Li, D., & Calderone, R. (2017) · Virulence · doi:10.1080/21505594.2016.1188235 · Cited by 67 · see citing works
Pappas, P. G., Vazquez, J. A., Oren, I., Rahav, G., Aoun, M., Bulpa, P., Ben-Ami, R., Ferrer, R., Mccarty, T., Thompson, G. R., et al. (2023) · Journal of Antimicrobial Chemotherapy · doi:10.1093/jac/dkad256 · Cited by 65 · see citing works
Zhu, Y., Kilburn, S., Kapoor, M., Chaturvedi, S., Shaw, K. J., & Chaturvedi, V. (2020) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.01124-20 · Cited by 50 · see citing works
Hodges, M. R., Ople, E., Wedel, P., Shaw, K. J., Jakate, A., Kramer, W. G., Marle, S. V., van Hoogdalem, E.-J., & Tawadrous, M. (2023) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.01623-22 · Cited by 52 · see citing works
Calcineurin Inhibitors Synergize with Manogepix to Kill Diverse Human Fungal Pathogens
Liston, S. D., Whitesell, L., Kapoor, M., Shaw, K. J., & Cowen, L. E. (2022) · Journal of Fungi · doi:10.3390/jof8101102 · Cited by 12 · see citing works
Self-citation
- Authors
- McLellan, C. A., Whitesell, L., King, O. D., Lancaster, A. K., Mazitschek, R., & Lindquist, S.
- Affiliations
- Whitehead Institute for Biomedical Research (US)
- Boston Biomedical Research Institute (US)
- Massachusetts General Hospital (US)
- Corresponding author
- Lindquist, Susan
- Venue
- ACS Chemical Biology
- Publisher
- American Chemical Society (ACS)
- Published
- 2012-06-22
- Volume
- 7
- Issue
- 9
- Pages
- 1520-1528
- ISSN
- 1554-8929 (ISSN-L)
- 1554-8937
- OpenAlex
- W2184546388
- Cited by
- 115
- References
- 42
- Citations by year
- 2026: 8 · 2025: 9 · 2024: 11 · 2023: 10 · 2022: 9 · 2021: 8 · 2020: 8 · 2019: 10 · 2018: 10 · 2017: 10 · 2016: 9 · 2015: 5 · 2014: 4 · 2013: 3 · 2012: 1
- FWCI
- 2.87
- Type
- journal article
- Language
- en
- Open access
- green
- hdl.handle.net
- Licence
- cc-by-nc
- Locations
- ACS Chemical Biology (publishedVersion)
- PubMed (publishedVersion)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc)
- Funders
- Howard Hughes Medical Institute
- Topics
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Health Sciences › Medicine › Epidemiology › Fungal Infections and Studies
- Keywords
- glycosylphosphatidylinositol anchor biosynthesis, antifungal drug targets, endoplasmic reticulum stress, Candida albicans, host-pathogen interaction, filamentation, β-glucan, macrophage cytokine response, small molecule inhibitors
- MeSH terms
- Amino Acid Sequence, Animals, Antifungal Agents, Candida, Candidiasis, Cell Line, Enzyme Inhibitors, Fungi, Host-Parasite Interactions, Humans, Molecular Sequence Data, Mycoses, Saccharomyces cerevisiae, Glycosylphosphatidylinositols, Saccharomyces cerevisiae Proteins, Mice
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
