Project description:Investigation of whole genome gene expression level changes in Candida glabrata CBS138 delta-vph2 mutant, compared to the wild-type strain in SC broth (pH5.0 and pH7.4). VPH2 gene encodes a protein that is the assembly factor of a functional V-ATPase. Loss of Vph2p leads to loss of a functional V-ATPase enzyme complex.
Project description:To delineate the interaction of Candida glabrata with host immune cells, we performed genome-wide transcriptional profiling analysis on THP-1 macrophage-internalized wild-type and chromatin remodeling defective mutant (Cgrsc3-a∆ and Cgrtt109∆) yeasts. Genes implicated in ergosterol biosynthesis, and high-affinity iron uptake and homeostasis were found to be down-regulated in C. glabrata wild-type and mutant cells upon macrophage internalization. Additionally, global gene expression profiles of RPMI-grown and macrophage-ingested Cgrsc3-a∆ and Cgrtt109∆ cells revealed down-regulation of genes involved in mitochondrial respiration under normal growth conditions and induction of genes required for generation of precursors of metabolites and energy upon macrophage internalization. To examine the behavior of Candida glabrata wild-type and chromatin remodeling defective mutants upon internalization by differentiated human monocytic THP-1 cells, we compared the transcript profiles of 10 hour RPMI-grown with those of 10 hour THP-1 macrophage internalized C. glabrata cells. Additionally, early transcriptional response of C. glabrata wild-type cells to macrophage internal milieu was examined post 2 hour THP-1 macrophage infection.
Project description:To understand the host response of C. glabrata, we have employed the whole genome microarray expression profiling to identify the differentially regulated genes. For this, differentiated THP-1 cells were infected with C. glabrata wt, Cgvps34Δ and Cgyps(1-11)Δ cells for 6 h. Uninfected THP-1 cells, treated with PBS were taken as control. Total RNA was extracted using TriZol Reagent followed by ethonol precipitation. The experiments were performed in biological duplicate.
