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A. fumigatus wild type and delta phoB(PHO80) strains transcriptome analysis for growth upon minimal medium 0.1 mM Pi

ABSTRACT: Phosphate is an essential ion for fungal growth, required for proper DNA and phospholipids biosyntesis, energetic metabolism and signal transduction. The systems for inorganic phosphate (Pi) acquisition in eukaryotic cells (PHO) have been characterized as a low-affinity (that assures a supply of Pi at normal or high external Pi concentrations) and a high-affinity (activated in response to Pi starvation). Here, as an initial step to understand the PHO pathway in A. fumigatus, we characterized the PHO80 homologue, phoB(PHO80). We showed that the delta phoB(PHO80) mutant has a severe polar growth defect and there is an interaction between PhoB(PHO80), calcineurin and calcium metabolism. Microarray hybridizations carried out with RNA obtained from wild type and delta phoB(PHO80) mutant cells showed that Afu4g03610 [phoD(PHO84)], Afu7g06350 [phoE9(PHO89)], Afu4g06020 [phoCPHO81)], and Afu2g09040 (vacuolar transporter Vtc4) were more expressed either in the delta phoB(PHO80) mutant background or in 0.1 mM Pi. Keywords: gene expression array-based (log2 ratio) Overall design: For the microarray experiments, 1.0 x 10(10) conidia of A. fumigatus wild type [delta akuB(ku80)] and delta phoB(PHO80) strain were used to inoculate 300 ml of liquid cultures (Minimal Medium with 0.1 mmol/L or 11 mmol/L of phosphate) in 500-ml erlenmeyer flasks and allow to grow for 6 and 10 hours in a reciprocal shaker (250 rpm) at 37°C. After this time, the cultures were centrifuged and their RNA were extracted. Hybridization experiments were competitive using probes derived from wild type and deltaphoB(PHO80) strain grown for 6 and 10 hours in MM with 0.1 mmol/L or 11 mmol/L of phosphate, using the wild type strain as the reference RNA for every hybridizantion. Normalized signal intensities were used to generate relative hybridization ratios (query/reference). Following normalization, the values for each gene's in-slide replicates were condensed (median variance <0.01), and corresponding flip-dye replicates were averaged to compensate for dye-specific effects (see Supplementary files linked below).

INSTRUMENT(S): TIGR PFGRC Aspergillus fumigatus v2 array designed primarily based on strain Af293

SUBMITTER: Gustavo Henrique Goldman   

PROVIDER: GSE9548 | GEO | 2007-11-09



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