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yEvo: Experimental evolution in high school classrooms selects for novel mutations and epistatic interactions that impact clotrimazole resistance in S. cerevisiae

View ORCID ProfileM. Bryce Taylor, View ORCID ProfileRyan Skophammer, View ORCID ProfileAlexa R. Warwick, View ORCID ProfileJosephine M. Boyer, yEvo Students, Margaux Walson, View ORCID ProfileChristopher R. L. Large, View ORCID ProfileAngela Shang-Mei Hickey, View ORCID ProfilePaul A. Rowley, View ORCID ProfileMaitreya J. Dunham
doi: https://doi.org/10.1101/2021.05.02.442375
M. Bryce Taylor
*Department of Genome Sciences, University of Washington, Seattle, WA 98195
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Ryan Skophammer
†Westridge School, Pasadena, CA 91105
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Alexa R. Warwick
‡Department of Fisheries and Wildlife, Michigan State University, East Lansing, MI 48824·
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Josephine M. Boyer
§Department of Biological Sciences, University of Idaho, Moscow, ID 83844
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yEvo Students
†Westridge School, Pasadena, CA 91105
**Moscow High School, Moscow, ID 83843
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Margaux Walson
*Department of Genome Sciences, University of Washington, Seattle, WA 98195
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Christopher R. L. Large
*Department of Genome Sciences, University of Washington, Seattle, WA 98195
††UW Molecular and Cellular Biology Program, University of Washington, Seattle, Washington 98195
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Angela Shang-Mei Hickey
*Department of Genome Sciences, University of Washington, Seattle, WA 98195
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Paul A. Rowley
§Department of Biological Sciences, University of Idaho, Moscow, ID 83844
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  • For correspondence: maitreya@uw.edu prowley@uidaho.edu
Maitreya J. Dunham
*Department of Genome Sciences, University of Washington, Seattle, WA 98195
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  • ORCID record for Maitreya J. Dunham
  • For correspondence: maitreya@uw.edu prowley@uidaho.edu
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Abstract

Antifungal resistance in pathogenic fungi is a growing global health concern. Non-pathogenic laboratory strains of Saccharomyces cerevisiae are a useful model for studying mechanisms of antifungal resistance that are relevant to understanding the same processes in pathogenic fungi. We developed a series of lab modules in which high school students used experimental evolution to study antifungal resistance by isolating azole-resistant S. cerevisiae and examining the genetic basis of resistance. All 99 sequenced clones from these experiments possessed mutations previously shown to impact azole resistance, demonstrating the efficacy of our protocols. We additionally found recurrent mutations in an mRNA degradation pathway and an uncharacterized mitochondrial protein (Csf1) that have possible mechanistic connections to azole resistance. The scale of replication in this high school-led initiative allowed us to identify epistatic interactions, as evidenced by pairs of mutations that occur in the same clone more frequently than expected by chance (positive epistasis) or less frequently (negative epistasis). We validated one of these pairs, a negative epistatic interaction between gain-of-function mutations in the multidrug resistance transcription factors Pdr1 and Pdr3. This high school-university collaboration can serve as a model for involving members of the broader public in the scientific process to make meaningful discoveries in biomedical research.

Competing Interest Statement

The authors have declared no competing interest.

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Posted May 03, 2021.
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yEvo: Experimental evolution in high school classrooms selects for novel mutations and epistatic interactions that impact clotrimazole resistance in S. cerevisiae
M. Bryce Taylor, Ryan Skophammer, Alexa R. Warwick, Josephine M. Boyer, yEvo Students, Margaux Walson, Christopher R. L. Large, Angela Shang-Mei Hickey, Paul A. Rowley, Maitreya J. Dunham
bioRxiv 2021.05.02.442375; doi: https://doi.org/10.1101/2021.05.02.442375
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yEvo: Experimental evolution in high school classrooms selects for novel mutations and epistatic interactions that impact clotrimazole resistance in S. cerevisiae
M. Bryce Taylor, Ryan Skophammer, Alexa R. Warwick, Josephine M. Boyer, yEvo Students, Margaux Walson, Christopher R. L. Large, Angela Shang-Mei Hickey, Paul A. Rowley, Maitreya J. Dunham
bioRxiv 2021.05.02.442375; doi: https://doi.org/10.1101/2021.05.02.442375

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