Project description:Transcriptome analysis of wild type, Hat1-deficient, Cac2-deficient and Rtt109-deficient Candida albicans cells prior to and after treatment with hydrogen was done to determine the transcriptional changes in these three mutatns in logarithmically growing cells as well as under oxidative stress conditions.
Project description:Candida albicans is a ubiquitous opportunistic pathogen that is the most prevalent cause of hospital-acquired fungal infections. In mammalian hosts, C. albicans is engulfed by phagocytes that attack the pathogen with DNA-damaging reactive oxygen species (ROS). Acetylation of histone H3 lysine 56 (H3K56) by the fungal-specific histone acetyltransferase Rtt109 is important for yeast model organisms to survive DNA damage and maintain genome integrity. To assess the importance of Rtt109 for C. albicans pathogenicity, we deleted the predicted homologue of Rtt109 in the clinical C. albicans isolate, SC5314. C. albicans rtt109 -/- mutant cells lack acetylated H3K56 (H3K56ac) and are hypersensitive to genotoxic agents. Additionally, rtt109 -/- mutant cells constitutively display increased H2A S129 phosphorylation and elevated DNA repair gene expression, consistent with endogenous DNA damage.
Project description:Candida albicans is a ubiquitous opportunistic pathogen that is the most prevalent cause of hospital-acquired fungal infections. In mammalian hosts, C. albicans is engulfed by phagocytes that attack the pathogen with DNA-damaging reactive oxygen species (ROS). Acetylation of histone H3 lysine 56 (H3K56) by the fungal-specific histone acetyltransferase Rtt109 is important for yeast model organisms to survive DNA damage and maintain genome integrity. To assess the importance of Rtt109 for C. albicans pathogenicity, we deleted the predicted homologue of Rtt109 in the clinical C. albicans isolate, SC5314. C. albicans rtt109 -/- mutant cells lack acetylated H3K56 (H3K56ac) and are hypersensitive to genotoxic agents. Additionally, rtt109 -/- mutant cells constitutively display increased H2A S129 phosphorylation and elevated DNA repair gene expression, consistent with endogenous DNA damage. Four independent pairs of biological replicate samples were analyzed to compare RNA levels in wild-type and rtt109 -/- cells. Two experiments were performed: first, in unperturbed cells grown in YPD and second, in cells exposed to H2O2. To account for dye effects, two of the four samples for each experiment were analyzed using Cy3 labeling of the wild-type sample and Cy5 labeling of the mutant sample; the dyes were swapped for the other two biological replicates.
Project description:ATAC-seq analysis of Candida albicans SC5314 treated or not treated with hydrogen peroxide to assess the accessible chromatin landscape upon oxidative stress in comparison to normal growth in YPD at 30 °C.
Project description:Candida albicans is a fungal opportunistic pathogen responsible for cause severe infections in immunocompromised patients. Despite nowadays an effective antifungal armoury is available, the emergence of antifungal resistance is promoting the urge to develop alternative treatments to combat this fungal pathogen. For this purpose it is essential to go deepen in the knowledge of C. albicans response to several stressors. With this aim we performed a proteomic study of C. albicans exposed to 5 and 10 mM of hydrogen peroxide and 40 and 60 mM of acetic acid taking advantage of data independent acquisition mass spectrometry. Four biological replicates of each condition were analysed. For protein identification we used a C. albicans spectral library previously constructed by DDA analysis allowing the DIA quantitation of nearby 2000 proteins. Changes in protein abundance after the treatments reflected highly different proteomic patterns after each treatment. The exposure of C. albicans to hydrogen peroxide led to an increase in many proteins related to oxidative stress response, proteasome and protein folding while few proteins from mitochondria decreased their abundance. In contrast, acetic acid treatment triggered the decrease in the abundance of proteins related to aminoacid biosynthesis, actin polymerization, oxidative stress response and proteasome. This extensive proteomic study provided valuable information for the development of new drug targets against decisive proteins for cell survival.
Project description:Transcriptional profiling of Candida albicans cells comparing control untreated C. albicans cells with sulfite-treated C. albicans cells. Sulfite is a toxic molecule that C. albicans encounters in its human host. Both wild type and ∆zcf2 mutant cells were used. The goal was to determine the effects of sulfite on C. albicans gene expression, and to determine which of the genes areZcf2-depedent.
Project description:RNA sequencing was performed on Candida albicans wild type cells (JC50) grown to exponential phase on YPD , YPD plus Nitrosative Stress 2.5mM DPTA NONOate, and compared to exponential Candida albicans hog1 deletion mutant cells grown on on YPD , YPD plus Nitrosative Stress 2.5mM DPTA NONOate. Three independent experiments were performed.
Project description:Candida albicans is an opportunistic human fungal pathogen. It exists as a member of human normal flora but can cause infection in immunocompromised individuals. Transition to pathogenic C. albicans requires a change in gene expression of various genes. As the histone-modifying enzymes can regulate gene expression, they can be considered as a factor controlling the virulence of C. albicans. Among them, it has been reported that the absence of H3K4 methyltransferase Set1 reduces the virulence of C. albicans. However, Set1-regulated genes responsible for this attenuated virulence phenotype are unknown. Here, we identified that the Set1 positively regulates the expression of mitochondrial protein genes and oxidative stress response-related genes through the methylation of H3K4. Levels of cellular mitochondrial reactive oxygen species (ROS) in Δset1 are also higher than those in the wild-type. Furthermore, Set1 deletion increases sensitivity to hydrogen peroxide (H2O2). Set1 deleted mutant also does not form its colony properly when it interacts with macrophage in vitro, in agreement with its attenuated virulence in vivo. Therefore, Set1 is required to inhibit cellular ROS production by positive regulation of mitochondrial protein genes and oxidative stress response-related genes expression, and consequently, the Set1-mediated gene expression assists C. albicans in defending against ROS generated from the host more rapidly.
Project description:The fungal pathogen Candida albicans produces dark-pigmented melanin when grown in a basal medium containing 1 mM l-DOPA as melanin substrate. In the widely used C. albicans strain SC5314, melanin appeared after 3-4 days of incubation in l-DOPA medium. The experiment was designed to reveal cadidate genes associated with melanin biosynthesis by expression profiling at different times of growth with and without L-DOPA added to the medium. Expression profiling of C. albicans revealed very few genes significantly up- or down-regulated by growth in l-DOPA.