Yanjun Song1, Shuixiu Li1, Yajing Zhao1, Yishan Zhang1, Yan Lv1, Yuanying Jiang2, Yan Wang2, Dongmei Li3, Hong Zhang4. 1. The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China; Institute of Mycology, Jinan University, Guangzhou, Guangdong, China. 2. New Drug Research and Development Center, School of Pharmacy, Second Military Medical University, Shanghai, China. 3. Department of Microbiology and Immunology, Georgetown University Medical Center, Washington, DC, USA. 4. The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China; Institute of Mycology, Jinan University, Guangzhou, Guangdong, China. Electronic address: tzhangh@jnu.edu.cn.
Abstract
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.
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 (infectedmice, 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.
Authors: Raquel Alonso-Roman; Antonia Last; Mohammad H Mirhakkak; Jakob L Sprague; Lars Möller; Peter Großmann; Katja Graf; Rena Gratz; Selene Mogavero; Slavena Vylkova; Gianni Panagiotou; Sascha Schäuble; Bernhard Hube; Mark S Gresnigt Journal: Nat Commun Date: 2022-06-09 Impact factor: 17.694
Authors: Giampiero Ferraguti; Sergio Terracina; Carla Petrella; Antonio Greco; Antonio Minni; Marco Lucarelli; Enzo Agostinelli; Massimo Ralli; Marco de Vincentiis; Giammarco Raponi; Antonella Polimeni; Mauro Ceccanti; Brunella Caronti; Maria Grazia Di Certo; Christian Barbato; Alessandro Mattia; Luigi Tarani; Marco Fiore Journal: Antioxidants (Basel) Date: 2022-01-11