Chemical Genetics Screen of EVP4593 Sensitivity in Budding Yeast Identifies Effects on Mitochondrial Structure and Function.
Chemical Genetics Screen of EVP4593 Sensitivity in Budding Yeast Identifies Effects on Mitochondrial Structure and Function.
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DOI：
10.17912/micropub.biology.000806
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发表时间：
2023
影响因子：
--
通讯作者：
Kerscher, Oliver
中科院分区：
文献类型：
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作者：
Hiestand, Lexie;Shen, Stella;Sloan, Willough;Nasiri, Hamid;Lashley, Dana;Kerscher, Oliver
线粒体是真核生物的重要细胞器。线粒体功能障碍可导致线粒体肌病，并可能导致神经退行性疾病、癌症和糖尿病。EVP 4593是一种具有治疗潜力的6-氨基喹唑啉衍生物，已被证明可抑制线粒体电子传递链的NADH-泛醌氧化还原酶（复合物I），导致活性氧（ROS）释放和ATP合成减少。在分离的线粒体中，EVP 4593在纳摩尔范围内抑制呼吸（IC 50 = 14-25 nM）。然而，还描述了对生物过程的其他EVP 4593特异性作用。与芽殖酵母中对线粒体功能的影响一致，我们发现当野生型细胞在不可发酵的碳源上生长时，EVP 4593 [>25μM]诱导明显的生长缺陷。这种对EVP 4593的敏感性因PDR 5的缺失而加剧，PDR 5是一种赋予多药耐药性的ABC转运蛋白。为了更好地了解EVP 4593影响的细胞途径和过程，我们对酵母敲除收集物进行了全基因组化学遗传学筛选。目的是鉴定当暴露于亚致死浓度的EVP 4593 [15µM]时表现出生长缺陷的酵母基因缺失菌株。我们的筛选确定了21个酵母基因，这些基因是在含甘油培养基中对15μM EVP 4593产生抗性所必需的。在我们的筛选中鉴定的基因在功能上涉及几个不同的类别，包括线粒体结构和功能，翻译调节和营养传感，细胞应激反应和解毒。此外，我们鉴定了与暴露于EVP 4593相关的细胞表型，包括线粒体结构的变化。总之，我们的研究代表了酵母中第一个全基因组筛选，以确定EVP 4593抗性所涉及的遗传途径和细胞保护机制，并揭示了这种小分子抑制剂影响线粒体结构和功能。
Mitochondria are essential eukaryotic organelles. Mitochondrial dysfunction can lead to mitochondrial myopathies and may contribute to neurodegenerative diseases, cancer, and diabetes. EVP4593, a 6-aminoquinazoline derivative with therapeutic potential, has been shown to inhibit NADH–ubiquinone oxidoreductase (Complex I) of the mitochondrial electron transport chain, causing the release of reactive oxygen species (ROS) and a reduction in ATP synthesis. In isolated mitochondria, EVP4593 inhibits respiration in the nanomolar range (IC 50 = 14-25 nM). However, other EVP4593-specific effects on biological processes have also been described. Consistent with an effect on mitochondrial function in budding yeast, we find that EVP4593 [>25µM] induces a pronounced growth defect when wildtype cells are grown on a non-fermentable carbon source. This sensitivity to EVP4593 is exacerbated by deletion of PDR5 , an ABC transporter that confers multidrug resistance. To better understand the cellular pathways and processes affected by EVP4593, we conducted a genome-wide chemical genetics screen of the yeast knockout collection. The objective was to identify yeast gene deletion strains that exhibit growth defects when subjected to a sublethal concentration of EVP4593 [15µM]. Our screen identified 21 yeast genes that are required for resistance to 15µM EVP4593 in glycerol-containing media. The genes identified in our screen are functionally involved in several distinct categories including mitochondrial structure and function, translational regulation and nutritional sensing, cellular stress response and detoxification. Additionally, we identified cellular phenotypes associated with the exposure to EVP4593, including changes in mitochondrial structure. In conclusion, our study represents the first genome-wide screen in yeast to identify the genetic pathways and cell-protective mechanisms involved in EVP4593 resistance and reveals that this small molecule inhibitor affects both mitochondrial structure and function.