Project description:Inhibition of fungal growth by Congo Red (CR) has been known for a long time and claimed to be associated to a specific binding to fibrillar chains of β-1,3-glucans which blocks cell wall polysaccharide synthesis. It has also been suggested that the effect of CR could be pleiotropic, especially because of its oxido-reductive properties. One way to elucidate the mechanism of action of CR is to identify transcription factors (TF) which regulate the response to CR and interrogate their regulon. The analysis of TF mutants with differential sensitivity to CR would be also a way to identify new regulatory pathways involved in cell wall biosynthesis. During the investigation of the sensitivity to CR of a TF mutant library, several mutants either sensitive or resistant to CR were analysed . The morphology of the conidial germination was affected differently by CR in the parental strain and TF mutants. Abnormal distorted swollen conidia called Quasimodo cells were seen in presence of the CR. The size of the Quasimodo cells in the resistant mutants are bigger and adsorb more CR than the germinating conidia of the sensitive or parental strain. This is associated to a difference in permeability of the mutants. The clearing of the drug from the medium by the resistant strain was mainly responsible for the survival and later growth of some of resting conidia of these mutants which did not adsorb intracellularly the drug whereas in the sensitive mutants all conidia adsorbed the drug and were consequently killed. Although there are some modifications of the composition of the cell wall, a transcriptomic analysis of the mutant showed that CR affected many different pathways, including iron and calcium metabolism. Accordingly, the susceptibility to other cell wall drugs of the sensitive and the resistant TF mutants varied with the mutants and not always in connexion with their sensitivity to CR. Interestingly, a new paradoxical effect of the CR was discovered in the resistant mutant since the growth of one of them could be stimulated after recovering from the CR stress.
Project description:We did transcription profiling on the effect of rlm1 (MAPK Slt2 transcription factor) deletion and swi3 (component of SWI/SNF complex involved in chromatin remodeling) deletion in genes involved in cell wall stress (Congo Red) response. Three biological samples were analyzed for each condition, and a microarray experiment were carried out for each sample 1. absence of the drug 2. presence of 30 µg/ml of Congo Red
Project description:We did transcription profiling on the effect of rlm1 (MAPK Slt2 transcription factor) deletion and swi3 (component of SWI/SNF complex involved in chromatin remodeling) deletion in genes involved in cell wall stress (Congo Red) response.
Project description:S. cerevisiae was grown on YEPD. For Congo Red experiments, yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h 30min. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with Congo Red to a final concentration of 30 μg/ml. At this time cells were collected t=0h. Keywords: repeat sample
Project description:S. cerevisiae rlm1Δ strain was grown on YEPD. For Congo Red experiments, yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h 30min. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with Congo Red to a final concentration of 30 μg/ml. Cells were collected at 4 hours of growth, frozen at -80 °C and processed for RNA extraction. Keywords: repeat sample
Project description:S. cerevisiae slt2Δ strain was grown on YEPD. For Congo Red experiments, yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h 30min. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with Congo Red to a final concentration of 30 μg/ml. Cells were collected at 4 hours of growth, frozen at -80 °C and processed for RNA extraction. Keywords: repeat sample
Project description:S. cerevisiae was grown on YEPD. For Congo Red experiments, yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h 30min. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with Congo Red to a final concentration of 30 μg/ml. Cells were collected at 6 hours of growth, frozen at -80 °C and processed for RNA extraction. Keywords: repeat sample
Project description:S. cerevisiae was grown on YEPD. For Congo Red experiments, yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h 30min. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with Congo Red to a final concentration of 30 μg/ml. Cells were collected at 4 hours of growth, frozen at -80 °C and processed for RNA extraction. Keywords: repeat sample
Project description:S. cerevisiae was grown on YEPD. For Congo Red experiments, yest cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h 30min. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with Congo Red to a final concentration of 30 μg/ml. Cells were collected at 2hours of growth, frozen at -80 °C and processed for RNA extraction. Keywords: repeat sample
Project description:S. cerevisiae was grown on YEPD. For Congo Red experiments, yest cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with Congo Red to a final concentration of 30 µg/ml. Cells were collected at 4hours of growth, frozen at -80 °C and processed for RNA extraction.