High-Throughput Screen for Identifying Small Molecules That Target Fungal Zinc Homeostasis
Open accessAbstract
Resistance to traditional antifungal drugs has increased significantly over the past three decades, making identification of novel antifungal agents and new targets an emerging priority. Based on the extraordinary zinc requirement of several fungal pathogens and their well-established sensitivity to zinc deprivation, we developed an efficient cell-based screen to identify new antifungal drugs that target the zinc homeostasis machinery. The screen is based on the zinc-regulated transcription factor Zap1 of Saccharomyces cerevisiae, which regulates transcription of genes like the high-affinity zinc transporter ZRT1. We generated a genetically modified strain of S. cerevisae that reports intracellular zinc deficiency by placing the coding sequence of green fluorescent protein (GFP) under the control of the Zap1-regulated ZRT1 promoter. After showing that the GFP fluorescence signal correlates with low intracellular zinc concentrations in this strain, a protocol was developed for screening small-molecule libraries for compounds that induce Zap1-dependent GFP expression. Comparison of control compounds and known modulators of metal metabolism from the library reveals a robust screen (Z' = 0.74) and validates this approach to the discovery of new classes of antifungal compounds that interfere with the intracellular zinc homeostasis. Given that growth of many pathogenic organisms is significantly impaired by zinc limitation; these results identify new types of antifungal drugs that target critical nutrient acquisition pathways.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
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?
Other FDA-approved medications, like atovaquone, target plasmodial cytochrome bc1 safely, which validates cytochrome bc1 as a suitable drug target.
“other FDA-approved medications, like atovaquone, target plasmodial cytochrome bc1 safely, which validates cytochrome bc1 as a suitable drug target [64].”
From Lansoprazole interferes with fungal respiration and acts synergistically with amphotericin B against multidrug-resistant <i>Candida auris</i> (2024) · cites this paper as [64]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Molecular Cloning: A Laboratory Manual
Sambrook, J., Fritsch, E., & Maniatis, T. (1989) · UCLA - Biblioteca de Ciencias de la Salud · Cited by 133,589 · see citing works
Molecular cloning: A laboratory manual
Deininger, P. L. (1990) · Analytical Biochemistry · doi:10.1016/0003-2697(90)90595-z · Cited by 84,684 · see citing works
Zhang, J.-H., Chung, T. D. Y., & Oldenburg, K. R. (1999) · SLAS Discovery · doi:10.1177/108705719900400206 · Cited by 6,847 · see citing works
Schiestl, R. H., & Gietz, R. D. (1989) · Current Genetics · doi:10.1007/bf00340712 · Cited by 2,143 · see citing works
Femtomolar Sensitivity of Metalloregulatory Proteins Controlling Zinc Homeostasis
Outten, C. E., & O'Halloran, A. T. V. (2001) · Science · doi:10.1126/science.1060331 · Cited by 1,489 · see citing works
Clinical, Cellular, and Molecular Factors That Contribute to Antifungal Drug Resistance
White, T. C., Marr, K. A., & Bowden, R. A. (1998) · Clinical Microbiology Reviews · doi:10.1128/cmr.11.2.382 · Cited by 1,353 · see citing works
The changing face of candidemia: emergence of non-Candida albicans species and antifungal resistance
Hong Nguyen, M., Peacock, J. E., Morris, A. J., Tanner, D. C., Nguyen, M. L., Snydman, D. R., Wagener, M. M., Rinaldi, M. G., & Yu, V. L. (1996) · The American Journal of Medicine · doi:10.1016/s0002-9343(95)00010-0 · Cited by 758 · see citing works
Resistance to echinocandin-class antifungal drugs.
Perlin, D. S. (2007) · Drug Resistance Updates · doi:10.1016/j.drup.2007.04.002 · Cited by 512 · see citing works
Taki, M., Wolford, J. L., & O’Halloran, T. V. (2003) · Journal of the American Chemical Society · doi:10.1021/ja039073j · Cited by 512 · see citing works
Self-citationMeasurement of Intracellular Free Zinc in Living Cortical Neurons: Routes of Entry
Sensi, S. L., Canzoniero, L. M. T., Yu, S. P., Ying, H. S.-C., Koh, J., Kerchner, G. A., & Choi, D. W. (1997) · Journal of Neuroscience · doi:10.1523/jneurosci.17-24-09554.1997 · Cited by 465 · see citing works
Applications of the green fluorescent protein in cell biology and biotechnology
Misteli, T., & Spector, D. L. (1997) · Nature Biotechnology · doi:10.1038/nbt1097-961 · Cited by 357 · see citing works
Fahrni, C. J., & O’Halloran, T. V. (1999) · Journal of the American Chemical Society · doi:10.1021/ja992709f · Cited by 332 · see citing works
Self-citationGenetically encoded biosensors based on engineered fluorescent proteins
Frommer, W. B., Davidson, M. W., & Campbell, R. E. (2009) · Chemical Society Reviews · doi:10.1039/b907749a · Cited by 321 · see citing works
Zhao, H. F., & Eide, D. J. (1997) · Molecular and Cellular Biology · doi:10.1128/mcb.17.9.5044 · Cited by 277 · see citing works
A new reporter cell line to monitor HIV infection and drug susceptibility in vitro
Gervaix, A., West, D., Leoni, L. M., Richman, D. D., Wong‐Staal, F., & Corbeil, J. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.9.4653 · Cited by 256 · see citing works
Ammendola, S., Pasquali, P., Pistoia, C., Petrucci, P., Petrarca, P., Rotilio, G., & Battistoni, A. (2007) · Infection and Immunity · doi:10.1128/iai.00559-07 · Cited by 248 · see citing works
Echinocandins: The newest class of antifungals
Sucher, A. J., Chahine, E. B., & Balcer, H. E. (2009) · Annals of Pharmacotherapy · doi:10.1345/aph.1m237 · Cited by 213 · see citing works
Zhao, H. F., Butler, E., Rodgers, J., Spizzo, T., Duesterhoeft, S., & Eide, D. J. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.44.28713 · Cited by 180 · see citing works
Homeostatic and Adaptive Responses to Zinc Deficiency in Saccharomyces cerevisiae
Eide, D. J. (2009) · Journal of Biological Chemistry · doi:10.1074/jbc.r900014200 · Cited by 148 · see citing works
Moreno, M. Á., Ibrahim‐Granet, O., Vicentefranqueira, R., Amich, J., Ave, P., Leal, F., Latgé, J., & Calera, J. A. (2007) · Molecular Microbiology · doi:10.1111/j.1365-2958.2007.05726.x · Cited by 135 · see citing works
Germ tube growth of Candida albicans.
Gow, N. A. R. (1997) · PubMed · Cited by 130 · see citing works
Saccharomyces cerevisiae Vacuole in Zinc Storage and Intracellular Zinc Distribution
Simm, C., Lahner, B., Salt, D. E., LeFurgey, A., Ingram, P., Yandell, B. S., & Eide, D. J. (2007) · Eukaryotic Cell · doi:10.1128/ec.00077-07 · Cited by 111 · see citing works
Self-citationEstrogen Decreases Zinc Transporter 3 Expression and Synaptic Vesicle Zinc Levels in Mouse Brain
Lee, J., Kim, J.-H., Hong, S. H., Lee, J. Y., Cherny, R. A., Bush, A. I., Palmiter, R. D., & Koh, J. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m309730200 · Cited by 98 · see citing works
Wu, C.-Y., Bird, A. J., Chung, L. M., Newton, M. A., Winge, D. R., & Eide, D. J. (2008) · BMC Genomics · doi:10.1186/1471-2164-9-370 · Cited by 97 · see citing works
Kim, S., Watanabe, K., Shirahata, T., & Watarai, M. (2004) · Journal of Veterinary Medical Science · doi:10.1292/jvms.66.1059 · Cited by 92 · see citing works
Vicentefranqueira, R., Moreno, M. A., Leal, F., & Calera, J. A. (2005) · Eukaryotic Cell · doi:10.1128/ec.4.5.837-848.2005 · Cited by 80 · see citing works
Winters, M. S., Chan, Q., Caruso, J. A., & Deepe, G. S. (2010) · The Journal of Infectious Diseases · doi:10.1086/656191 · Cited by 76 · see citing works
Kim, M. J., Kil, M., Jung, J., & Kim, J. (2008) · PubMed · Cited by 76 · see citing works
Weston, B. F., Brenot, A., & Caparon, M. G. (2009) · Infection and Immunity · doi:10.1128/iai.01299-08 · Cited by 63 · see citing works
Control of Dimorphism in Candida Albicans by Zinc: Effect on Cell Morphology and Composition
YAMAGUCHI, H. (1975) · Journal of General Microbiology · doi:10.1099/00221287-86-2-370 · Cited by 62 · see citing works
Fungal susceptibility to zinc deprivation
Lulloff, S. J., Hahn, B. L., & Sohnle, P. G. (2004) · Journal of Laboratory and Clinical Medicine · doi:10.1016/j.lab.2004.07.007 · Cited by 61 · see citing works
A model for the germ tube formation and mycelial growth form ofCandida albicans
Gow, N. A. R., & Gooday, G. W. (1984) · Medical Mycology · doi:10.1080/00362178485380211 · Cited by 61 · see citing works
Lamore, S. D., & Wondrak, G. T. (2011) · BioMetals · doi:10.1007/s10534-011-9441-6 · Cited by 48 · see citing works
The Metal Chelating and Chaperoning Effects of Clioquinol: Insights from Yeast Studies
Li, C., Wang, J., & Zhou, B. (2010) · Journal of Alzheimer s Disease · doi:10.3233/jad-2010-100024 · Cited by 46 · see citing works
Sexual maturation and reproductive zinc physiology in the female squirrelfish
Thompson, E. D., Mayer, G. D., Walsh, P. J., & Högstrand, C. (2002) · Journal of Experimental Biology · doi:10.1242/jeb.205.21.3367 · Cited by 44 · see citing works
Identification and characterization of a high-affinity zinc uptake system in Neisseria gonorrhoeae
Chen, C., & Morse, S. A. (2001) · FEMS Microbiology Letters · doi:10.1111/j.1574-6968.2001.tb10781.x · Cited by 39 · see citing works
Zhu, M., & Fahl, W. E. (2000) · Analytical Biochemistry · doi:10.1006/abio.2000.4875 · Cited by 34 · see citing works
Antifungals: Need to search for a new molecular target
Sangamwar, A. T., Deshpande, U., & Pekamwar, S. S. (2008) · Indian Journal of Pharmaceutical Sciences · doi:10.4103/0250-474x.44588 · Cited by 30 · see citing works
Mondal, R., Koev, G., Pilot-Matias, T., et al. (2009) · Antiviral Research · doi:10.1016/j.antiviral.2008.12.012 · Cited by 16 · see citing works
Ismail, A., & Bedell, G. W. (1986) · Mycopathologia · doi:10.1007/bf00437261 · Cited by 6 · see citing works
Preemptive therapy in Candida infections
Biliri, K., & Michael, D. (2009) · Reviews in Medical Microbiology · doi:10.1097/mrm.0b013e3283341397 · Cited by 1 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Zinc and Iron Homeostasis: Target-Based Drug Screening as New Route for Antifungal Drug Development
Simm, C., & May, R. C. (2019) · Frontiers in Cellular and Infection Microbiology · doi:10.3389/fcimb.2019.00181 · Cited by 26 · see citing works
Self-citationSalama, E. A., Elgammal, Y., Wijeratne, A., Lanman, N. A., Utturkar, S. M., Farhangian, A., Li, J., Meunier, B., Hazbun, T. R., & Seleem, M. N. (2024) · Emerging Microbes & Infections · doi:10.1080/22221751.2024.2322649 · Cited by 16 · see citing works
- Authors
- Simm, C., Luan, C.-H., Weiss, E., & O'Halloran, T.
- Affiliations
- Northwestern University (US)
- Corresponding author
- Simm, Claudia
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2011-09-29
- Volume
- 6
- Issue
- 9
- Pages
- e25136
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC3182986
- OpenAlex
- W2152713010
- Cited by
- 34
- References
- 41
- Citations by year
- 2025: 2 · 2024: 1 · 2023: 2 · 2022: 3 · 2021: 1 · 2020: 2 · 2019: 2 · 2018: 2 · 2017: 2 · 2016: 3 · 2015: 6 · 2014: 2 · 2013: 3 · 2012: 2
- FWCI
- 0.91
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Figshare (submittedVersion, cc-by)
- Funders
- National Aeronautics and Space Administration (NNA04CC36G)
- Northwestern University
- National Institutes of Health (GM038784)
- National Institute of General Medical Sciences (R37GM038784, R01GM038784)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Health Sciences › Nursing › Nutrition and Dietetics › Trace Elements in Health
- Keywords
- antifungal drug discovery, Saccharomyces cerevisiae, zinc deprivation, high-throughput screening, small molecule library, metal metabolism, antifungal resistance, nutrient acquisition
- MeSH terms
- Antifungal Agents, Flow Cytometry, Homeostasis, Saccharomyces cerevisiae, Transcription Factors, Zinc, Cation Transport Proteins, Saccharomyces cerevisiae Proteins, Green Fluorescent Proteins
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
