Hiestand Lexie, Shen Stella, Sloan Willough, Nasiri Hamid, Lashley Dana, Kerscher Oliver
Biology, William & Mary, Williamsburg, Virginia, United States.
Chemistry, William & Mary, Williamsburg, Virginia, United States.
MicroPubl Biol. 2023 Apr 25;2023. doi: 10.17912/micropub.biology.000806. eCollection 2023.
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 = 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 , 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.
线粒体是真核生物必不可少的细胞器。线粒体功能障碍可导致线粒体肌病，并可能引发神经退行性疾病、癌症和糖尿病。EVP4593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中的NADH-泛醌氧化还原酶（复合体I），导致活性氧（ROS）释放并减少ATP合成。在分离的线粒体中，EVP4593在纳摩尔范围内抑制呼吸作用（IC = 14 - 25 nM）。然而，也有其他关于EVP4593对生物过程的特异性作用的描述。与对出芽酵母线粒体功能的影响一致，我们发现当野生型细胞在非发酵碳源上生长时，EVP4593 [>25µM] 会导致明显的生长缺陷。缺失赋予多药耐药性的ABC转运蛋白会加剧对EVP4593的这种敏感性。为了更好地理解受EVP4593影响的细胞途径和过程，我们对酵母基因敲除文库进行了全基因组化学遗传学筛选。目的是鉴定在亚致死浓度的EVP4593 [15µM] 作用下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 EVP4593产生抗性所必需的。我们筛选中鉴定出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、翻译调控和营养感知、细胞应激反应和解毒。此外，我们确定了与接触EVP4593相关的细胞表型，包括线粒体结构的变化。总之，我们的研究代表了在酵母中首次进行的全基因组筛选，以确定参与EVP4593抗性的遗传途径和细胞保护机制，并揭示这种小分子抑制剂会影响线粒体的结构和功能。
