Project description:Transcript profiling was performed using a wild-type C. albicans strain (CaI8+CIp10). Cells were cultured in 50 ml SC-pH3.0 to a cell density of 1x10^7 cells per ml and then either treated with control (0 mM acetic acid) or stress inducing (20 mM acetic acid) doses. Cells from the same culture were harvested after 300 min treatment. Three independent biological replicates were obtained for each condition.
Project description:Based on studies in S. pombe (Chen et al., 2003), we predicted that conditions which activate C. albicans Hog1 would result in the induction of a common set of genes that are regulated by this SAPK (Smith et al., 2004). Hence in this study we compared the global transcriptional responses of wild-type and hog1 C. albicans cells to environmental stresses that are known to activate the Hog1 SAPK. We compared the homozygous hog1/hog1 null mutant (JC50) with the isogenic HOG1 reintegrant (hog1/hog1/HOG1: JC52) because this controlled for any secondary mutations that might have been introduced during the construction of the null mutant. We have shown that this reintegrant is indistinguishable from its parental wild-type strain RM1000 (HOG1/HOG1) with respect to their stress phenotypes (Smith et al., 2004) and their expression of stress genes (Supplementary Data). Three conditions were chosen for transcript profiling: osmotic stress imposed by 0.3 M NaCl, oxidative stress imposed by 5 mM H2O2, and heavy metal stress imposed by 0.5 mM CdSO4. Each of these treatments stimulates the phosphorylation and nuclear accumulation of this Hog1 SAPK within a 10-min time frame (Smith et al., 2004). Furthermore, significant differences in stress regulated gene expression are observed within this time scale (Enjalbert et al., 2003; Smith et al., 2004). Hence, we analyzed the C. albicans transcriptome after a 10-min exposure to each stress condition. Although some stress genes might be missed by analyzing a single time point, most C. albicans stress genes are induced within 10 min (Enjalbert et al., 2003). At least four independent biological replicates were analyzed for each condition
Project description:We investigated how Candida albicans adapts its transcriptional pattern to a sudden availability of glucose in the extracellular medium, having previously grown on a non-fermentable carbon source.
Project description:Candida albicans wild type strain BWP17 and mutant hac1 were submitted either to DTT or tunicamycin, both agents being able to trigger the Unfolded Protein Response.
Project description:Wild type Candida albicans (CAI8 containing CIp10) cultures were grown in triplicate at 1 x 107 cells ml-1 in 50 ml SC-pH3.0 broth. Cultures were then treated with 0, 20, 120 or 300<br>mM acetic acid. Cells were harvested after zero, 10, 30, 60, 120, 210 and 300 min treatment. RNA isolated from triplicate biological replicates were compared with a standard pooled control (untreated cells).