yEvo: experimental evolution in high school classrooms selects for novel mutations that impact clotrimazole resistance in Saccharomyces cerevisiae.
yEvo: experimental evolution in high school classrooms selects for novel mutations that impact clotrimazole resistance in Saccharomyces cerevisiae.
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DOI：
10.1093/g3journal/jkac246
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发表时间：
2022-11-04
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影响因子：
--
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中科院分区：
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病原真菌的抗真菌耐药性是一个日益增长的全球健康问题。酿酒酵母的非致病性实验室菌株是研究抗真菌药物抗性机制的重要模型，这些机制与了解致病真菌中的相同过程有关。我们已经开发了一系列的实验室模块，高中学生使用实验进化，通过分离耐唑的S。酿酒酵母突变体和检查抗性的遗传基础。我们已经对这些实验中的99个克隆进行了测序，发现所有这些克隆都具有先前显示的影响唑类耐药性的突变，从而验证了我们的方法。我们还发现了mRNA降解途径和未表征的线粒体蛋白（Csf1）中的复发性突变，这些突变可能与唑类耐药有关。在这一举措中的复制规模使我们能够确定候选上位相互作用，如在同一克隆中发生的突变对比偶然（正上位性）或不太频繁（负上位性）的预期更频繁所证明的。我们验证了其中一对，在多药耐药转录因子Pdr1和Pdr3的功能获得性突变之间的负上位相互作用。这种高中与大学的合作可以作为一种模式，让更广泛的公众参与科学过程，在生物医学研究中做出有意义的发现。我们从一个独特的合作教育研究奋进我们称之为yEvo，其中学生使用酵母的实验进化来研究抗真菌药物耐药性的遗传基础，日益增长的公共卫生和农业问题的数据。除了在已知的唑类耐药基因中发现突变外，我们还发现了新的基因，并将其中一些基因与可能的机制联系起来。我们还提供了突变之间上位相互作用的证据，这表明以前没有与这种表型相关的途径之间的联系。
Antifungal resistance in pathogenic fungi is a growing global health concern. Nonpathogenic laboratory strains of Saccharomyces cerevisiae are an important model for studying mechanisms of antifungal resistance that are relevant to understanding the same processes in pathogenic fungi. We have developed a series of laboratory modules in which high school students used experimental evolution to study antifungal resistance by isolating azole-resistant S. cerevisiae mutants and examining the genetic basis of resistance. We have sequenced 99 clones from these experiments and found that all possessed mutations previously shown to impact azole resistance, validating our approach. 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 initiative allowed us to identify candidate 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. We present data from a unique collaborative education-research endeavor we call yEvo, in which students use experimental evolution of yeast to study the genetic basis of antifungal resistance, a growing public health and agricultural problem. In addition to finding mutations in known azole resistance genes, we discovered new genes and tie some of these to possible mechanisms. We additionally provide evidence for epistatic interactions between mutations, which suggest connections between pathways that have not previously been linked to this phenotype.
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影响因子：
14.9
作者：
Dubreuil B;Sass E;Nadav Y;Heidenreich M;Georgeson JM;Weill U;Duan Y;Meurer M;Schuldiner M;Knop M;Levy ED
通讯作者：
Levy ED
影响因子：
3.3
作者：
Copic, Alenka;Dorrington, Mariana;Miller, Elizabeth A.
通讯作者：
Miller, Elizabeth A.
影响因子：
3.2
作者：
Cowen, LE;Sanglard, D;Kohn, LM
通讯作者：
Kohn, LM
影响因子：
6.4
作者：
Carolus H;Pierson S;Muñoz JF;Subotić A;Cruz RB;Cuomo CA;Van Dijck P
通讯作者：
Van Dijck P
DOI：
10.1073/pnas.1702314114
发表时间：
2017-08-01
影响因子：
11.1
作者：
Buskirk, Sean W.;Peace, Ryan Emily;Lang, Gregory I.
通讯作者：
Lang, Gregory I.