title: Dual action antifungal small molecule modulates multidrug efflux and TOR signaling.
abstract: There is an urgent need for new strategies to treat invasive fungal infections, which are a leading cause of human mortality. Here, we establish two activities of the natural product beauvericin, which potentiates the activity of the most widely deployed class of antifungal against the leading human fungal pathogens, blocks the emergence of drug resistance, and renders antifungal-resistant pathogens responsive to treatment in mammalian infection models. Harnessing genome sequencing of beauvericin-resistant mutants, affinity purification of a biotinylated beauvericin analog, and biochemical and genetic assays reveals that beauvericin blocks multidrug efflux and inhibits the global regulator TORC1 kinase, thereby activating the protein kinase CK2 and inhibiting the molecular chaperone Hsp90. Substitutions in the multidrug transporter Pdr5 that enable beauvericin efflux impair antifungal efflux, thereby impeding resistance to the drug combination. Thus, dual targeting of multidrug efflux and TOR signaling provides a powerful, broadly effective therapeutic strategy for treating fungal infectious disease that evades resistance.
authors: Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
authors: Tanvi
authors: T
authors: Shekhar-Guturja
authors: Natural Products Center, School of Natural Resources and the Environment, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona, USA.
authors: G M Kamal B
authors: GM
authors: Gunaherath
authors: Natural Products Center, School of Natural Resources and the Environment, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona, USA.
authors: E M Kithsiri
authors: EM
authors: Wijeratne
authors: Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors: Jean-Philippe
authors: JP
authors: Lambert
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, USA.
authors: Anna F
authors: AF
authors: Averette
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, USA.
authors: Soo Chan
authors: SC
authors: Lee
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, USA.
authors: Taeyup
authors: T
authors: Kim
authors: Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea.
authors: Yong-Sun
authors: 0000-0001-9573-5752
authors: YS
authors: Bahn
authors: Department of Biotechnology and Biosciences, University of Milano-Bicocca and SYSBIO, Centre of Systems Biology, Milan, Italy.
authors: Farida
authors: F
authors: Tripodi
authors: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada.
authors: Ron
authors: R
authors: Ammar
authors: Institute of Biochemistry, Heinrich Heine University Duesseldorf, Duesseldorf, Germany.
authors: Katja
authors: K
authors: Döhl
authors: Department of Molecular Biology, University of Geneva, Geneva, Switzerland.
authors: Karolina
authors: K
authors: Niewola-Staszkowska
authors: Institute of Biochemistry, Heinrich Heine University Duesseldorf, Duesseldorf, Germany.
authors: Lutz
authors: L
authors: Schmitt
authors: Department of Molecular Biology, University of Geneva, Geneva, Switzerland.
authors: Robbie J
authors: RJ
authors: Loewith
authors: Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.|Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors: Frederick P
authors: FP
authors: Roth
authors: Institute of Microbiology, University Hospital Lausanne and University Hospital Center, Lausanne, Switzerland.
authors: Dominique
authors: D
authors: Sanglard
authors: Department of Medicine, University of Wisconsin, Madison, Wisconsin, USA.|Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, Wisconsin, USA.
authors: David
authors: D
authors: Andes
authors: Faculty of Pharmaceutical Sciences, The University of British Columbia, Vancouver, British Columbia, Canada.
authors: Corey
authors: 0000-0002-4016-8874
authors: C
authors: Nislow
authors: Department of Biotechnology and Biosciences, University of Milano-Bicocca and SYSBIO, Centre of Systems Biology, Milan, Italy.
authors: Paola
authors: 0000-0001-5898-5883
authors: P
authors: Coccetti
authors: Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.|Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors: Anne-Claude
authors: AC
authors: Gingras
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, USA.
authors: Joseph
authors: J
authors: Heitman
authors: Natural Products Center, School of Natural Resources and the Environment, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona, USA.
authors: A A Leslie
authors: AA
authors: Gunatilaka
authors: Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
authors: Leah E
authors: LE
authors: Cowen
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology*; D047630:Depsipeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D025141:Drug Resistance, Fungal / Q000187:drug effects; D018432:Drug Resistance, Multiple / Q000187:drug effects; D005658:Fungi / Q000187:drug effects* / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D009181:Mycoses / Q000188:drug therapy / Q000382:microbiology; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents; D047630:Depsipeptides; D018841:HSP90 Heat-Shock Proteins; C092438:PDR5 protein, S cerevisiae; D047428:Protein Kinase Inhibitors; D029701:Saccharomyces cerevisiae Proteins; D054852:Small Molecule Libraries; C004456:beauvericin; D058570:TOR Serine-Threonine Kinases