Transcription profiling time series of Candida albicans treated with different doses of acetic acid
ABSTRACT: 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 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).
MNL1, the Candida albicans homologue of an orphan Msn2-like gene (YER130c in Saccharomyces cerevisiae) has no known function. Here we report that MNL1 regulates weak acid stress responses. Deletion of MNL1 prevents the long-term adaptation of C. albicans cells to weak acid stresses and compromises their global transcriptional response under these conditions. The promoters of Mnl1-dependent genes contain a novel STRE-like element (SLE) that imposes Mnl1-dependent, weak acid stress-induced transcr ...[more]
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:C. albicans (strain CAI4-CIp10) was grown according to CRISP SOP. An overnight culture was started from a single colony in YPD-Tris,(100mM Tris.HCl) pH 7.4 and incubated overnight at 30C in shaking incubator. The next day, 500 ul culture was inoculated into 50 ml YPD-Tris, pH 7.4. The following day a fresh culture was inoculated in YPD-Tris to an OD600 of 0.2 and grown to OD600 of 0.8 at 30C in a shaking incubator. At this point the culture was split, diluted back to OD600 of 0.2 in fresh medium and stress agents added at time =0 (XS = 5mM H2O2, OS = 1M NaCl or a combination, OSXS). After 10 min the cultures were harvested by centrifugation and cell pellets flash frozen in liquid nitrogen. Three independent cultures were grown for RNA extraction for each isolate at each condition.<br><br>The control sample was obtained from cells with no stress agents added, harvested at time=0. RNA was extracted and transcript profiling carried out by microarray analysis using custom microarrays (Eurogentec).
Project description:A pre-culture in NGY (overnight 30°C) was used to inoculate RPMI-1640 medium to an initial optical density at 600 nm (OD600) of 0.05. Cells were then grown to mid exponential phase (OD600 = 0.50.6) at the appropriate temperature (25°C or 35°C) before the culture was<br><br>split into two aliquots and fluconazole (32 ?g ml1 final concentration) added to one; the other serving as the control. After 2 h the cells were harvested by centrifugation and flash<br><br>frozen in liquid nitrogen. Three independent cultures were grown for RNA extraction for each isolate at each condition. RNA was prepared from cell pellets with TRIZOL® reagent (Invitrogen) and a mechanical disruption method.
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.