Objective

Alcohol dehydrogenase I is encoded by ADH1 in Candida albicans, and is one of the key enzymes in fungal metabolism by which it catalyzes the conversion from acetaldehyde to ethanol. The role of the associated protein Adh1p, encoded by ADH1 in fungal pathogenicity has not been thoroughly studied despite its near ubiquity in the fungal kingdom. Using C. albicans as a model, this study proposes to determine the possible pathogenic roles for ADH1 and its possible underlying mechanisms.

Methods

The SAT1 flipper strategy was used to construct the ADH1 deletion mutant. Growth curves and spot assay were used to compare growth and cell viability of the mutant to wild type C. albicans. Three host model systems (infected mice, C. elegans, and G. mellonella) were used to investigate the effects of ADH1 deletion in vivo on C. albicans pathogenicity. Then, adhesion, hyphal formation, biofilm formation, cell surface hydrophobicity (CSH) and RT-qPCR were performed to investigate the effects of ADH1 deletion in vitro on C. albicans virulence. Finally, Xfe 96 seahorse assay, ROS level, mitochondrial membrane potential, and intracellular ATP content were used to determine the effects of ADH1 deletion on bioenergetics.

Results

ADH1 deletion has no effects on the growth and cell viability of C. albicans, but significantly prolongs survival time in each of the three host models, decreases fungal burden in kidney and liver, and lessens pathological tissue damage (P < 0.05). In addition, ADH1 deletion significantly increases CSH and reduces C. albicans virulence in terms of adhesion, hyphal formation and biofilm formation in accord with the downregulation of virulence-related genes such as ALS1, ALS3, HWP1, and CSH1 (P < 0.05). For bioenergetics, ADH1 deletion has no obvious effect on glycolysis, but a lack of ADH1 significantly increases ROS levels and decreases mitochondrial membrane potential and intracellular ATP content even through the mitochondrial oxygen consumption rate and NADH/NAD⁺ ratio are elevated (P < 0.05).

Conclusion

Our results suggest that the fermentative enzyme ADH1 is required for the pathogenicity of C. albicans under one of the presumed mechanisms viaits effects on oxidative phosphorylation activities in mitochondria.

中文翻译：

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ADH1通过刺激氧化磷酸化促进白色念珠菌的致病性。

客观的

酒精脱氢酶I在白色念珠菌中由ADH1编码，是真菌代谢中的关键酶之一，通过它催化乙醛向乙醇的转化。尽管由ADH1编码的相关蛋白Adh1p在真菌致病性中的作用尚未得到充分研究，但在真菌界却普遍存在。以白色念珠菌为模型，本研究建议确定ADH1的可能致病作用及其可能的潜在机制。

方法

该SAT1脚蹼策略用于构建ADH1缺失突变体。用生长曲线和斑点检测法比较突变体对野生型白色念珠菌的生长和细胞活力。三个主机模型系统（感染的小鼠，线虫（C.elegans） ，和蜡螟）被用来研究的影响ADH1缺失体内对白色念珠菌的致病性。然后，进行粘附，菌丝形成，生物膜形成，细胞表面疏水性（CSH）和RT-qPCR来研究体外ADH1缺失对白色念珠菌的影响。毒力。最后，使用Xfe 96海马测定法，ROS水平，线粒体膜电位和细胞内ATP含量来确定ADH1缺失对生物能学的影响。

结果

ADH1缺失对白色念珠菌的生长和细胞活力没有影响，但是在三个宿主模型中的每一个中均显着延长了存活时间，减少了肾脏和肝脏的真菌负担，并减轻了病理组织损伤（P  <0.05）。此外，ADDH1缺失与ALS1，ALS3，HWP1和CSH1等与毒力相关的基因的下调一致，在粘附，菌丝形成和生物膜形成方面显着增加了CSH并降低了白色念珠菌的毒力（P  <0.05）。对于生物能学，ADH1缺失对糖酵解没有明显影响，但缺乏ADH1即使提高了线粒体的耗氧率和NADH / NAD ⁺比，也显着提高了ROS水平并降低了线粒体膜电位和细胞内ATP含量（P  <0.05）。

结论

我们的研究结果表明，发酵酶ADH1是通过其对线粒体氧化磷酸化活性的影响，在一种假想的念珠菌的致病性中所必需的。