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Dual action antifungal small molecule modulates multidrug efflux and TOR signaling
Open accessAbstract
There is an urgent need for new strategies to treat invasive fungal infections, which are a leading cause of human mortality. 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 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 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 fungal infectious disease that evades resistance.
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TL;DR
Beauvericin potentiates azole antifungals against major fungal pathogens by dual inhibition of multidrug efflux and TORC1 signaling, leading to CK2 activation and Hsp90 impairment. This combination blocks resistance emergence and shows therapeutic benefit in mammalian infection models.
Key findings
- Beauvericin had no antifungal activity alone but enhanced azole efficacy against *S. cerevisiae*, *C. albicans*, *C. neoformans*, and *A. fumigatus*; against *C. albicans*, fluconazole–beauvericin synergy gave a fractional inhibitory concentration index (FICI) of <0.1 and converted fungistatic azole activity into a fungicidal combination.
- Beauvericin blocked emergence of azole resistance: plating 1×10⁵ *C. albicans* cells on 32 μg/ml fluconazole with 20 μg/ml beauvericin prevented resistant colonies.
- Beauvericin-resistant mutants selected at 100 μg/ml beauvericin in transporter-deficient backgrounds had mutations in *PDR5* (five mutants) or *CKB1*/*CKB2* (ten mutants, six independent events); Pdr5 substitutions enabled beauvericin efflux while impairing azole efflux.
- Affinity purification with biotinylated beauvericin identified 12 proteins at FDR ≤1%, including Cka2 and Tor2; beauvericin inhibited TORC1-dependent Sch9 phosphorylation, activated CK2, impaired Hsp90 clients, and in mice infected with 5×10⁶ *C. albicans*, beauvericin (4 mg/kg) plus fluconazole (0.5 mg/kg) gave potent therapeutic benefit; in a rat catheter biofilm model, the combination sterilized catheters.
Methods
- Drug combination and resistance assays used azole Etest strips, checkerboard matrices, broth microdilution MIC testing, and spotting assays with *S. cerevisiae*, *C. albicans*, *C. neoformans*, and *A. fumigatus*, including *C. albicans* clinical isolates.
- Target identification combined selection of beauvericin-resistant mutants in ABC-transporter-deficient backgrounds, whole-genome sequencing, Sanger validation, and affinity purification coupled to mass spectrometry with a biotinylated beauvericin analog, analyzed by SAINTexpress.
- Mechanistic assays included radiolabeled [³H]-fluconazole accumulation, rhodamine-6G transport and accumulation, Pdr5 ATPase activity, CK2 kinase assays with Sic1 phosphorylation, qRT-PCR, Sch9/Ypk1 and Hog1 western blots, β-galactosidase reporters, and murine/rat infection models.
Limitations
- The paper does not state a dedicated limitations section.
- Evident from the text: the ABC16 selection experiment was performed once, and the murine disseminated-infection model used no blinding.
- Some target conclusions are indirect: excess unmodified beauvericin did not compete the biotinylated analog from most proteins, including Cka2, and beauvericin did not alter recombinant human CK2 activity, leaving fungal-specific or indirect effects unresolved.
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33 Own result · 14 Own interpretation · 13 Own method · 11 Not checked yet · 1 Partly supported · accuracy not yet measured
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In the paper
“inhibits the global regulator TORC1 kinase”
- Also stated in: § Beauvericin activates CK2 and inhibits TORC1 · plain
“Beauvericin caused a significant reduction in Sch9 phosphorylation and a modest reduction in Ypk1 phosphorylation (Fig. 4e, Supplementary Fig. 10).”
- Also stated in: § Discussion · plain
“beauvericin inhibits TORC1”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
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- Also stated in: § Beauvericin activates CK2 and inhibits TORC1 · plain
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In the paper
“Whole genome sequencing of ten resistant mutants revealed that each harboured a mutation in either CKB1 or CKB2, encompassing six independent mutational events (Fig. 3a).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
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“We used a dose response matrix (checkerboard) to corroborate the synergy between the azole fluconazole and beauvericin against C. albicans, with a fractional inhibitory concentration index (FICI) of <0.1 (Fig. 1b).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole efficacy and blocks resistance
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In the paper
“Beauvericin reduced resistance of all isolates (Fig. 1c), with cidality of the combination evident at higher azole concentrations”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole efficacy and blocks resistance
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“Inclusion of beauvericin in the medium at a concentration that did not impair growth (20 μg/ml) blocked the emergence of resistance (Fig. 1d).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole efficacy and blocks resistance
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“deletion of either CKA1 or CKA2 from the ABC16 strain confers partial resistance to beauvericin (Supplementary Fig. 6)”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
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“Transferring cells from the checkerboard onto drug-free rich medium revealed that beauvericin transforms azole fungistatic activity into a fungicidal combination (Fig. 1b).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole efficacy and blocks resistance
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“Beauvericin increased Sic1 phosphorylation, suggesting that it activates CK2 (Fig. 4b and Supplementary Fig. 8a).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin activates CK2 and inhibits TORC1
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“Tor2 was identified in all three experiments”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Biotinylated beauvericin interacts with Cka2 and Tor2
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In the paper
“potentiates the activity of the most widely deployed class of antifungal against the leading human fungal pathogens”
- Also stated in: § Beauvericin enhances azole efficacy and blocks resistance · plain
“Beauvericin had no antifungal activity alone, but enhanced azole efficacy against the model yeast Saccharomyces cerevisiae and fungal pathogens C. albicans, C. neoformans, and A. fumigatus (Fig. 1a).”
- Also stated in: § Introduction · plain
“Beauvericin enhances azole efficacy against the leading human fungal pathogens, Candida albicans, Aspergillus fumigatus, and Cryptococcus neoformans”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
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- Rests on an assumption: The mechanism of beauvericin action (inhibition of Pdr5, TORC1, CK2, and Hsp90) identified in S. cerevisiae and C. albicans is conserved in other human fungal pathogens such as A. fumigatus and C. neoformans.
- Also stated in: § Beauvericin enhances azole efficacy and blocks resistance · plain
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“beauvericin blocks multidrug efflux”
- Also stated in: § Beauvericin enhances azole intracellular accumulation · plain
“Beauvericin inhibited Pdr5 ATPase activity and transport of rhodamine-6G (Fig. 2e).”
- Also stated in: § Discussion · plain
“Beauvericin inhibits multidrug efflux”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
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- Also stated in: § Beauvericin enhances azole intracellular accumulation · plain
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In the paper
“Deletion of either CKB1 or CKB2 in the ABC16 background conferred full beauvericin resistance, suggesting that the mutations identified cause loss of function of CK2 subunits (Fig. 3b).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
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“Two experiments identified Cka2 as a top interacting protein, corroborating the link between beauvericin and CK2.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Biotinylated beauvericin interacts with Cka2 and Tor2
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“Expression of PDR5G538R as the only PDR5 allele in a yor1Δ mutant conferred full beauvericin resistance (Fig. 2b).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
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“Beauvericin had no antifungal activity alone”
- Also stated in: § Discussion · plain
“it has little antifungal activity alone due to efflux mediated by the ABC transporter Yor1”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole efficacy and blocks resistance
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“These are not loss-of-function mutations, as deleting PDR5 in the yor1Δ strain did not confer beauvericin resistance (Fig. 2a).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
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“Beauvericin is not a substrate of wild-type Pdr5, as deletion of PDR5 did not enhance beauvericin susceptibility (Fig. 2a,b).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
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“We screened S. cerevisiae strains each lacking one of 16 ATP-binding cassette (ABC) transporter genes14, and found that only the strain lacking YOR1 was sensitive to beauvericin (Supplementary Fig. 2).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
What the sources say
- Not checked yetcites [14] Suzuki, Y. et al. Knocking out multigene redundancies via cycles of sexual assortment and fluorescence selection. Nat. Methods 8, 159–164 (2011). source
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“Twelve proteins were identified in at least two of the three experiments (Fig. 4a).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Biotinylated beauvericin interacts with Cka2 and Tor2
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“Each of the resistant mutants had nonsynonymous mutations in the ABC transporter gene PDR5 (Fig. 2a), which mapped to transmembrane domains or proximal regions.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
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“We found that beauvericin caused increased fluconazole accumulation in S. cerevisiae and C. albicans (Fig. 2c, bottom panel).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole intracellular accumulation
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“Substitutions in the multidrug transporter Pdr5 that enable beauvericin efflux impair antifungal efflux”
- Also stated in: § Pdr5 substitutions enable beauvericin efflux · plain
“Representative mutants yor1Δ R1 and R5 show increased accumulation of radiolabelled azole (Fig. 2c, top panel).”
- Also stated in: § Discussion · plain
“substitutions in the ABC transporter Pdr5 that alter substrate specificity and enable beauvericin efflux concomitantly impair azole efflux”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
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“Activation of calcineurin in response to fluconazole was blocked by the calcineurin inhibitor FK506, and partially blocked by the Hsp90 inhibitor geldanamycin and by beauvericin (Fig. 5a).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin impairs Hsp90 function
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“As a combination, there was potent therapeutic benefit of beauvericin and fluconazole (Fig. 6a).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin potentiates azole therapeutic efficacy
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“We observed potent antagonism, such that beauvericin suppressed the antifungal activity of rapamycin (Fig. 4c).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin activates CK2 and inhibits TORC1
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“Addition of either CK2α or CK2α2β2 caused increased Sic1 phosphorylation that was not altered by beauvericin (Supplementary Fig. 8c).”
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“deletion of CKB1 or CKA2 reduced the impact of beauvericin on Hog1 activation (Fig. 5c).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin impairs Hsp90 function
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“As a drug combination, beauvericin and fluconazole sterilized the catheter (Fig. 6b,c).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin potentiates azole therapeutic efficacy
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“Beauvericin potently induced the HSR (Fig. 5b), consistent with a repressive effect on Hsp90 function.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin impairs Hsp90 function
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“beauvericin impaired Hog1 activation in the ABC16 strain (Fig. 5c, Supplementary Fig. 11).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin impairs Hsp90 function
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“This suggests that Yor1 effluxes beauvericin, and enables selection of resistant mutants.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
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“inhibiting the molecular chaperone Hsp90”
- Also stated in: § Beauvericin impairs Hsp90 function · hedged
“Beauvericin potently induced the HSR (Fig. 5b), consistent with a repressive effect on Hsp90 function.”
- Also stated in: § Discussion · plain
“beauvericin causes inhibition of Hsp90 function”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
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“Together, this suggests that Pdr5 substrate specificity is altered in the mutants, enabling beauvericin efflux at the expense of other substrates.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
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“suggesting that beauvericin exerts antifungal activity via CK2.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
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“Thus, loss of function of CK2 subunits confers beauvericin resistance, suggesting that beauvericin exerts antifungal activity via CK2.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
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“Thus, dual targeting of multidrug efflux and TOR signaling provides a powerful, broadly effective therapeutic strategy for fungal infectious disease that evades resistance.”
- Also stated in: § Introduction · boosted
“Thus, dual targeting of multidrug efflux and TOR signaling provides a powerful and broadly effective therapeutic strategy for fungal infectious disease.”
- Also stated in: § Discussion · boosted
“Our findings reveal that dual targeting of multidrug efflux and TOR signaling provides a powerful strategy to enhance antifungal efficacy, impede drug resistance, and render resistant pathogens responsive to treatment.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
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- Rests on an assumption: The mechanism of beauvericin action (inhibition of Pdr5, TORC1, CK2, and Hsp90) identified in S. cerevisiae and C. albicans is conserved in other human fungal pathogens such as A. fumigatus and C. neoformans.
- Rests on an assumption: The therapeutic benefit of the beauvericin-fluconazole combination in mammalian infection models is mediated by the same dual targeting of multidrug efflux and TOR signaling.
- Rests on an assumption: The Pdr5 substitutions that enable beauvericin efflux and impair azole efflux are representative of resistance mechanisms that would emerge in clinical settings.
- Also stated in: § Introduction · boosted
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Beauvericin must have an additional target beyond Pdr5 to mitigate antifungal activity in the absence of azoles, as Pdr5 is not required for viability of the wild type or yor1Δ mutant (Fig. 2).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“activating protein kinase CK2”
- Also stated in: § Beauvericin activates CK2 and inhibits TORC1 · hedged
“Beauvericin increased Sic1 phosphorylation, suggesting that it activates CK2 (Fig. 4b and Supplementary Fig. 8a).”
- Also stated in: § Discussion · plain
“stimulating protein kinase CK2”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Also stated in: § Beauvericin activates CK2 and inhibits TORC1 · hedged
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“thereby impeding resistance to the drug combination”
- Also stated in: § Pdr5 substitutions enable beauvericin efflux · plain
“Thus, mutations in PDR5 that enable beauvericin efflux impede azole efflux, thereby blocking the emergence of resistance to the drug combination.”
- Also stated in: § Discussion · plain
“thereby impeding the emergence of resistance to the drug combination.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Abstract
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: The Pdr5 substitutions that enable beauvericin efflux and impair azole efflux are representative of resistance mechanisms that would emerge in clinical settings.
- Also stated in: § Pdr5 substitutions enable beauvericin efflux · plain
None of the sources cited for this claim have been checked yet.
In the paper
“Hsp90 regulates drug resistance by stabilizing the protein phosphatase calcineurin9,10”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Introduction
What the sources say
- Not checked yetcites [9] Cowen, L.E. & Lindquist, S. Hsp90 potentiates the rapid evolution of new traits: drug resistance in diverse fungi. Science 309, 2185–2189 (2005). source
- Not checked yetcites [10] Singh, S.D. et al. Hsp90 governs echinocandin resistance in the pathogenic yeast Candida albicans via calcineurin. PLoS Pathog. 5, e1000532 (2009). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Thus, beauvericin exerts a repressive effect on Hsp90 via activation of CK2 signaling, providing a mechanism by which it abrogates azole resistance and creates a fungicidal drug combination (Fig. 5d).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin impairs Hsp90 function
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Thus, beauvericin has profound therapeutic benefits in combination with azoles in mammalian infection models.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin potentiates azole therapeutic efficacy
boosteddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: The therapeutic benefit of the beauvericin-fluconazole combination in mammalian infection models is mediated by the same dual targeting of multidrug efflux and TOR signaling.
In the paper
“TORC1 regulates phosphorylation of CK2 via the effector kinase Kns121.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Biotinylated beauvericin interacts with Cka2 and Tor2
What the sources say
- Not checked yetcites [21] Sanchez-Casalongue, M.E. et al. Differential phosphorylation of a regulatory subunit of protein kinase CK2 by target of rapamycin complex 1 signaling and the Cdc-like kinase Kns1. J. Biol. Chem. 290, 7221–7233 (2015). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Probably not a substantial claim. 20
- It may state two separate claims as one. 13
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Inhibition of Hsp90 abrogates drug resistance in diverse human fungal pathogens, rendering resistant infections responsive to treatment7–9.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Introduction
What the sources say
- Supportedcites [7] Cowen, L.E. et al. Harnessing Hsp90 function as a powerful, broadly effective therapeutic strategy for fungal infectious disease. Proc. Natl. Acad. Sci. USA 106, 2818–2823 (2009). source
The Abstract states that Hsp90 inhibition abrogates azole resistance in C. albicans and reduces echinocandin resistance in Aspergillus. The Results report that combining Hsp90 inhibitors with antifungals rescued larvae infected with a resistant C. albicans isolate and improved survival in larvae infected with A. fumigatus. These are host-model findings, not evidence of treatment success in human patients. (LLM, conf=0.92)
- Partly supportedcites [8] Robbins, N. et al. Hsp90 governs dispersion and drug resistance of fungal biofilms. PLoS Pathog. 7, e1002257 (2011). sourceExperimental — accuracy not yet measured
The Discussion reports that Hsp90 inhibition made fluconazole effective against C. albicans biofilms in a rat infection model and improved antifungal efficacy against A. fumigatus biofilms. It does not establish that resistance was abrogated or resistant infections became responsive across diverse human fungal pathogens; broader activity is presented as a possibility. (LLM, conf=0.83)
- Not checked yetcites [9] Cowen, L.E. & Lindquist, S. Hsp90 potentiates the rapid evolution of new traits: drug resistance in diverse fungi. Science 309, 2185–2189 (2005). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may state two separate claims as one.
- Supported
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Both the increase in fluconazole accumulation and the drug synergy were blocked in an S. cerevisiae strain lacking Pdr5 (Supplementary Fig. 5 and Fig. 2d).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole intracellular accumulation
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Beauvericin inhibited Pdr5 ATPase activity and transport of rhodamine-6G (Fig. 2e).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole intracellular accumulation
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“expression of wild-type CKB1 in the ABC16 ckb1Δ strain restored beauvericin susceptibility, while resistance was maintained upon expression of CKB1E220* (Fig. 3b).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
In the paper
“The interaction between beauvericin and rapamycin was abrogated by deletion of genes encoding FKBP12 (FPR1 in S. cerevisiae, RBP1 in C. albicans)25 and impervious to deletion of PDR5 (Supplementary Fig. 9), confirming the effects are mediated via TOR.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin activates CK2 and inhibits TORC1
What the sources say
- Not checked yetcites [25] Cruz, M.C. et al. Rapamycin and less immunosuppressive analogs are toxic to Candida albicans and Cryptococcus neoformans via FKBP12-dependent inhibition of TOR. Antimicrob. Agents Chemother. 45, 3162–3170 (2001). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Beauvericin caused a significant reduction in Sch9 phosphorylation and a modest reduction in Ypk1 phosphorylation (Fig. 4e, Supplementary Fig. 10).”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin activates CK2 and inhibits TORC1
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Thus, CK2 and TOR may be binding targets of beauvericin through which it exerts antifungal activity.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Biotinylated beauvericin interacts with Cka2 and Tor2
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Thus, beauvericin enhances azole efficacy through inhibition of Pdr5, thereby causing increased azole accumulation within the fungal cell.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin enhances azole intracellular accumulation
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“qRT-PCR reactions were performed in triplicate for biological duplicate experiments.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Quantitative reverse transcription (RT)-PCR
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Together, these results suggest that beauvericin activates CK2 by binding to Tor2 and inhibiting TORC1 function.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin activates CK2 and inhibits TORC1
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
In the paper
“it was found to be well tolerated by human cells and to potentiate azole efficacy against Candida parapsilosis in a systemic infection model by a mechanism that remains enigmatic12.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Introduction
What the sources say
- Not checked yetcites [12] Zhang, L. et al. High-throughput synergy screening identifies microbial metabolites as combination agents for the treatment of fungal infections. Proc. Natl. Acad. Sci. USA 104, 4606–4611 (2007). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Experiments were performed in biological duplicates; plates were incubated at 30°C for five days and photographed each day.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Selection experiments
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“No blinding was done.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Murine model of disseminated candidiasis
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“In these conditions, recovered cells were still in late log phase.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Radiolabelled fluconazole accumulation assay
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“The difference of both assays corresponds to the OM-sensitive ATPase activity of Pdr550.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > ATPase Activity Assay
What the sources say
- Not checked yetcites [50] Ernst, R. et al. A mutation of the H-loop selectively affects rhodamine transport by the yeast multidrug ABC transporter Pdr5. Proc. Natl. Acad. Sci. USA 105, 5069–5074 (2008). source
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“The average coverage of each genome was sufficient for confident variant detection and is listed in Supplementary Table 3.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Whole genome sequencing
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“AP-MS was performed in three independent experiments.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Affinity purification coupled to mass spectrometry with biotinylated beauvericin
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“CKB1 and CKB2 encode the non-redundant beta regulatory subunits of protein kinase CK2, which is an essential serine/threonine protein kinase that phosphorylates over 300 substrates, including Hsp9017.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Mutations in CK2 subunits confer beauvericin resistance
What the sources say
- Not checked yetcites [17] Miyata, Y. Protein kinase CK2 in health and disease: CK2: the kinase controlling the Hsp90 chaperone machinery. Cell. Mol. Life Sci. 66, 1840–1849 (2009). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
In the paper
“Pdr5 transports a diverse set of structurally unrelated compounds including azoles, cycloheximide, and rhodamines15.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Pdr5 substitutions enable beauvericin efflux
What the sources say
- Not checked yetcites [15] Prasad, R. & Goffeau, A. Yeast ATP-binding cassette transporters conferring multidrug resistance. Annu. Rev. Microbiol. 66, 39–63 (2012). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“All strains were assessed in biological triplicate experiments with technical duplicates.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Drug susceptibility and checkerboard assays
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“No growth defect was observed in comparison to media without methanol.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Selection experiments
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“In S. cerevisiae and C. albicans, CK2 phosphorylates Hsp90, thereby modulating chaperone function27,28.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin impairs Hsp90 function
What the sources say
- Not checked yetcites [27] Mollapour, M. et al. Threonine 22 phosphorylation attenuates Hsp90 interaction with cochaperones and affects its chaperone activity. Mol. Cell 41, 672–681 (2011). source
- Not checked yetcites [28] Diezmann, S., Michaut, M., Shapiro, R.S., Bader, G.D. & Cowen, L.E. Mapping the Hsp90 genetic interaction network in Candida albicans reveals environmental contingency and rewired circuitry. PLoS Genet. 8, e1002562 (2012). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“Hsp90 regulates cellular responses required to survive drug-induced stress, and inhibition of Hsp90 transforms azoles from fungistatic to fungicidal7,9.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Beauvericin impairs Hsp90 function
What the sources say
- Not checked yetcites [7] Cowen, L.E. et al. Harnessing Hsp90 function as a powerful, broadly effective therapeutic strategy for fungal infectious disease. Proc. Natl. Acad. Sci. USA 106, 2818–2823 (2009). source
- Not checked yetcites [9] Cowen, L.E. & Lindquist, S. Hsp90 potentiates the rapid evolution of new traits: drug resistance in diverse fungi. Science 309, 2185–2189 (2005). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“The selection experiment with the ABC16 strain was performed once, while the selection experiment with the yor1Δ strain was done in biological triplicates.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Selection experiments
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Control experiments were performed in which the biotinylated-beauvericin was omitted.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Affinity purification coupled to mass spectrometry with biotinylated beauvericin
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Mutations identified in CKB1, CKB2, and PDR5 were validated by Sanger sequencing.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Whole genome sequencing
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
In the paper
“Hsp90, an essential molecular chaperone that stabilizes diverse cellular regulators6.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Introduction
What the sources say
- Not checked yetcites [6] Leach, M.D., Klipp, E., Cowen, L.E. & Brown, A.J.P. Fungal Hsp90: a biological transistor that tunes cellular outputs to thermal inputs. Nat. Rev. Microbiol. 10, 693–704 (2012). source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Infected mice were randomly chosen for each group.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Murine model of disseminated candidiasis
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Survival data for all of the tested mice were included in the statistical analysis.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Murine model of disseminated candidiasis
plainn = 10- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
The paper relies on this without support.
The mechanism of beauvericin action (inhibition of Pdr5, TORC1, CK2, and Hsp90) identified in S. cerevisiae and C. albicans is conserved in other human fungal pathogens such as A. fumigatus and C. neoformans.
“potentiates the activity of the most widely deployed class of antifungal against the leading human fungal pathogens”
“Thus, dual targeting of multidrug efflux and TOR signaling provides a powerful, broadly effective therapeutic strategy for fungal infectious disease that evades resistance.”
The paper relies on this without support.
The therapeutic benefit of the beauvericin-fluconazole combination in mammalian infection models is mediated by the same dual targeting of multidrug efflux and TOR signaling.
“As a combination, there was potent therapeutic benefit of beauvericin and fluconazole (Fig. 6a).”
“Our findings reveal that dual targeting of multidrug efflux and TOR signaling provides a powerful strategy to enhance antifungal efficacy, impede drug resistance, and render resistant pathogens responsive to treatment.”
The paper relies on this without support.
The Pdr5 substitutions that enable beauvericin efflux and impair azole efflux are representative of resistance mechanisms that would emerge in clinical settings.
“Substitutions in the multidrug transporter Pdr5 that enable beauvericin efflux impair antifungal efflux, thereby impeding resistance to the drug combination.”
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Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“The natural product beauvericin, which potentiates azole activity via both inhibition of TOR and multidrug efflux, is effective against diverse fungal pathogens including C. albicans, C. neoformans, and A. fumigatus87.”
From Molecular mechanisms governing antifungal drug resistance (2023) · cites this paper as [87]
In the citing paper
“The natural product beauvericin, which potentiates azole activity via both inhibition of TOR and multidrug efflux, is effective against diverse fungal pathogens including C. albicans, C. neoformans, and A. fumigatus87.”
From Molecular mechanisms governing antifungal drug resistance (2023) · cites this paper as [87]
In the citing paper
“Combination therapy is an alternative approach to overcoming the growing microbial infection resistance [23,24].”
From Lansoprazole interferes with fungal respiration and acts synergistically with amphotericin B against multidrug-resistant Candida auris (2024) · cites this paper as [23]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Results > Biotinylated beauvericin interacts with Cka2 and Tor2
“AP-MS was performed under three distinct buffer conditions with biological duplicates and negative controls.”
§ Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling > Online Methods > Affinity purification coupled to mass spectrometry with biotinylated beauvericin
“Experiments 1 and 2 were performed using 0.1% NP-40 buffer, and experiment 3 was performed using 1% Triton X-100 buffer to allow for efficient extraction of membrane proteins.”
Citations
Cites
Sensitive detection of somatic point mutations in impure and heterogeneous cancer samples
Cibulskis, K., Lawrence, M. S., Carter, S. L., Sivachenko, A., Jaffe, D. B., Sougnez, C. L., Gabriel, S., Meyerson, M. M., Lander, E. S., & Getz, G. A. (2013) · Nature Biotechnology · doi:10.1038/nbt.2514 · Cited by 4,965
Hidden Killers: Human Fungal Infections
Brown, G. D., Denning, D. W., Gow, N. A. R., Levitz, S. M., Netea, M. G., & White, T. C. (2012) · Science Translational Medicine · doi:10.1126/scitranslmed.3004404 · Cited by 4,284
Targets for Cell Cycle Arrest by the Immunosuppressant Rapamycin in Yeast
Heitman, J., Movva, N. R., & Hall, M. N. (1991) · Science · doi:10.1126/science.1715094 · Cited by 2,010
Self-citationLoewith, R., Jacinto, E., Wullschleger, S., Lorberg, A., Crespo, J. L., Bonenfant, D., Oppliger, W., Jenoe, P., & Hall, M. N. (2002) · Molecular Cell · doi:10.1016/s1097-2765(02)00636-6 · Cited by 1,886
Self-citationSynergistic drug combinations tend to improve therapeutically relevant selectivity
Lehár, J., Krueger, A., Avery, W., Heilbut, A. M., Johansen, L. M., Price, E. R., Rickles, R. J., Short, G. F., Staunton, J. E., Jin, X., et al. (2009) · Nature Biotechnology · doi:10.1038/nbt.1549 · Cited by 959
The promise and peril of chemical probes
Arrowsmith, C. H., Audia, J. E., Austin, C., Baell, J., Bennett, J., Blagg, J., Bountra, C., Brennan, P. E., Brown, P. J., Bunnage, M. E., et al. (2015) · Nature Chemical Biology · doi:10.1038/nchembio.1867 · Cited by 839
SAINT: probabilistic scoring of affinity purification–mass spectrometry data
Choi, H., Larsen, B., Lin, Z.-Y., Breitkreutz, A., Mellacheruvu, D., Fermin, D., Qin, Z. S., Tyers, M., Gingras, A.-C., & Nesvizhskii, A. I. (2011) · Nature Methods · doi:10.1038/nmeth.1541 · Cited by 800
Self-citationSch9 Is a Major Target of TORC1 in Saccharomyces cerevisiae
Urban, J., Soulard, A., Huber, A., Lippman, S., Mukhopadhyay, D., Deloche, O., Wanke, V., Anrather, D., Ammerer, G., Riezman, H., et al. (2007) · Molecular Cell · doi:10.1016/j.molcel.2007.04.020 · Cited by 798
Self-citationHsp90 Potentiates the Rapid Evolution of New Traits: Drug Resistance in Diverse Fungi
Cowen, L. E., & Lindquist, S. (2005) · Science · doi:10.1126/science.1118370 · Cited by 698
Self-citationSAINTexpress: Improvements and additional features in Significance Analysis of INTeractome software
Teo, G., Liu, G., Zhang, J., Nesvizhskii, A. I., Gingras, A., & Choi, H. (2013) · Journal of Proteomics · doi:10.1016/j.jprot.2013.10.023 · Cited by 693
Self-citation
Cited by
Sorted by citations
Molecular Evolution of Antifungal Drug Resistance
Robbins, N., Caplan, T., & Cowen, L. E. (2017) · Annual Review of Microbiology · doi:10.1146/annurev-micro-030117-020345 · Cited by 441
Self-citationMolecular mechanisms governing antifungal drug resistance
Lee, Y., Robbins, N., & Cowen, L. E. (2023) · npj Antimicrobials and Resistance · doi:10.1038/s44259-023-00007-2 · Cited by 304
Self-citationPribis, J. P., Garcı́a-Villada, L., Zhai, Y., Lewin‐Epstein, O., Wang, A. Z., Liu, J., Xia, J., Mei, Q., Fitzgerald, D. M., Bos, J., et al. (2019) · Molecular Cell · doi:10.1016/j.molcel.2019.02.037 · Cited by 177
Geddes‐McAlister, J., & Shapiro, R. S. (2018) · Annals of the New York Academy of Sciences · doi:10.1111/nyas.13739 · Cited by 162
Advancements and challenges in antifungal therapeutic development
Puumala, E., Fallah, S., Robbins, N., & Cowen, L. E. (2024) · Clinical Microbiology Reviews · doi:10.1128/cmr.00142-23 · Cited by 128
Self-citation
Metadata
- Authors
- Shekhar-Guturja, T., Gunaherath, G. M. K. B., Wijeratne, E. M. K., Lambert, J.-P., Averette, A. F., Lee, S. C., Kim, T., Bahn, Y.-S., Tripodi, F., Ammar, R., Döhl, K., Niewola-Staszkowska, K., Schmitt, L., Loewith, R. J., Roth, F. P., Sanglard, D., Andes, D., Nislow, C., Coccetti, P., Gingras, A.-C., Heitman, J., Gunatilaka, A. A. L., & Cowen, L. E.
- Affiliations
- University of Arizona (US)
- Mount Sinai Hospital (CA)
- Duke Medical Center (US)
- Yonsei University (KR)
- University of Milano-Bicocca (IT)
- University of Toronto (CA)
- Heinrich Heine University Düsseldorf (DE)
- University of Geneva (CH)
- Centre Hospitalier Universitaire Vaudois (CH)
- University of Wisconsin–Madison (US)
- University of British Columbia (CA)
- Corresponding author
- Cowen, Leah E
- ORCID
- Gunaherath, G M Kamal B
- Wijeratne, E M Kithsiri
- Lambert, Jean-Philippe
- Averette, Anna F
- Lee, Soo Chan
- Kim, Taeyup
- Bahn, Yong-Sun
- Tripodi, Farida
- Ammar, Ron
- Schmitt, Lutz
- Loewith, Robbie J
- Roth, Frederick P
- Sanglard, Dominique
- Andes, David
- Nislow, Corey
- Coccetti, Paola
- Gingras, Anne-Claude
- Heitman, Joseph
- Gunatilaka, A A Leslie
- Cowen, Leah E
- Venue
- Nature Chemical Biology
- Published
- 2016-08-29
- Volume
- 12
- Issue
- 10
- Pages
- 867-875
- ISSN
- 1552-4450 (ISSN-L)
- 1552-4469
- PMC
- PMC5030160
- OpenAlex
- W2509038340
- Semantic Scholar
- 703e3c9f64b33ff7226b5d45e13b72dc9ef9243c
- Cited by
- 101
- References
- 57
- Citations by year
- 2026: 5 · 2025: 10 · 2024: 11 · 2023: 9 · 2022: 15 · 2021: 8 · 2020: 13 · 2019: 10 · 2018: 9 · 2017: 9 · 2016: 2
- FWCI
- 9.24
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Chemical Biology (publishedVersion)
- PubMed (publishedVersion)
- Archive ouverte UNIGE (University of Geneva) (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- SERVAL (Université de Lausanne) (acceptedVersion, other-oa)
- IRIS (submittedVersion)
- BOA (University of Milano-Bicocca) (submittedVersion)
- Archive ouverte UNIGE (University of Geneva) (submittedVersion, cc-by)
- Funders
- TD Bank
- Genome Canada
- Ministry of Trade, Industry and Energy (N0001720)
- University of Toronto
- National Institutes of Health (5r01ai112595-04)
- Canadian Institutes of Health Research (119520_1, MOP-86452)
- Natural Sciences and Engineering Research Council of Canada (462167, 06261)
- National Cancer Institute (R01CA090265)
- National Institute of General Medical Sciences (P41GM094060)
- National Institute of Allergy and Infectious Diseases (R01-AI112595, P01AI104533, R56AI112595)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › Microbial Natural Products and Biosynthesis
- Keywords
- beauvericin, antifungal resistance, multidrug efflux, TOR signaling, casein kinase 2, HSP90 inhibition, invasive fungal infections, natural products, drug repurposing
- MeSH terms
- Antifungal Agents, Fungi, Microbial Sensitivity Tests, Mycoses, Signal Transduction, Drug Resistance, Multiple, ATP-Binding Cassette Transporters, HSP90 Heat-Shock Proteins, Drug Resistance, Fungal, Saccharomyces cerevisiae Proteins, Protein Kinase Inhibitors, Depsipeptides, Small Molecule Libraries, TOR Serine-Threonine Kinases
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.
A language model drew the claims from the paper on 2026-10-09, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).
A second model labelled each claim on 2026-10-09 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).
Cited sources are checked with a language model’s help (gpt-6-sol).
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 ›
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