CaIPF7817 is involved in the regulation of redox homeostasis in Candida albicans
Section snippets
Materials and methods
Strains, culture, and growth of C. albicans. Candida albicans strains used in this study were listed in Table 1. The strains were cultured at 30 °C in YPD medium (1% Difco yeast extract, 2% Difco peptone, 2% dextrose) or YPD medium supplemented with 0.005% uridine (Sigma, USA). Agar (1.5%) was added to prepare the solid media. The URA3 transformants were selected on MIN (2% glucose, 0.67% Difco yeast nitrogen base without amino acids) agar plates supplemented with 25 μg/mL histidine. The ura3
Generation and characterization of ipf7817 mutants
To investigate the roles of CaIPF7817 in C. albicans, we disrupted the two alleles of CaIPF7817 sequentially in RM1000 strain using the URA-blaster method and 5-FOA selection, yielding the following strains: the Ura+ipf7817/IPF7817 strain (JH1U), Ura−ipf7817/IPF7817 strain (JH1), Ura+ipf7817/ipf7817 strain (JH2U), and Ura−ipf7817/ipf7817 strain (JH2) (Table 1). The strategy to disrupt both copies of CaIPF7817 was depicted in Fig. 1A. Southern blot analysis of DNA from the constructed strains
Discussion
In this work, we interrupted both copies of CaIPF7817 in C. albicans and studied the role of CaIPF7817 in cellular redox regulation. Intracellular ROS level, mitochondrial membrane potential, expressions of some important redox-related genes and GSH/GSSG ratio were measured to reveal changes in cellular redox condition after CaIPF7817 deletion. Without CaIPF7817, the intracellular ROS level was increased; mitochondrial membrane potential was elevated; the expressions of some important
Acknowledgments
This work was funded by a grant from the National Natural Science Foundation of China (30500628) and the National 973 Program of China (2005CB523105). We thank J. Pla for kindly providing us the C. albicans strain RM1000 and W.A. Fonzi for plasmid p5921.
References (28)
- et al.
Yeast signaling pathways in the oxidative stress response
Mutat. Res.
(2005) - et al.
Cap1p is involved in multiple pathways of oxidative stress response in Candida albicans
Free Radic. Bio. Med.
(2006) - et al.
Low dose (-)deprenyl is cytoprotective: it maintains mitochondrial membrane potential and eliminates oxygen radicals
Life Sci.
(2005) - et al.
Manganese activation of superoxide dismutase 2 in the mitochondria of Saccharomyces cerevisiae
J. Biol. Chem.
(2005) Changing patterns and trends in systemic fungal infections
J. Antimicrob. Chemother.
(2005)An overview of fungal infections
Drugs
(2001)- et al.
Immunosuppression and recovery of drug-impaired host resistance against Candida albicans infection by oxoglaucine
Pharmacol. Res.
(2000) Yeast stress responses
Oxidative stress responses of the yeast Saccharomyces cerevisiae
Yeast
(1998)- et al.
Redox control of exocytosis: regulatory role of NADPH, thioredoxin, and glutaredoxin
Diabetes
(2005)
Apoptosis induced by environmental stresses and amphotericin B in Candida albicans
Proc. Natl. Acad. Sci. USA
Regulation of redox homeostasis in the yeast Saccharomyces cerevisiae
Physiol. Plant
Free radicals in the physiological control of cell function
Physiol. Rev.
Proteomic analysis of the oxidative stress response in Candida albicans
Proteomics
Cited by (9)
Cap1p attenuates the apoptosis of Candida albicans
2013, FEBS Journal
- 1
These authors contributed equally to this work.