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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Comparison of transcript abundance in aerial mycelium of the Magnaporthe oryzae TRA1-deleted mutant and its parental strain

ABSTRACT: In order to understand the function of the Tra1p transcription factor we identified potentially Tra1p-regulated genes by comparison of transcript abundance in aerial mycelium of the tra1- mutant and the parental strain 70-15. Two-condition experiment, wild-type vs. TRA1-deleted mutant. 4 biological replicates with dye-swaps. Each microarry slide had 2 arrays with 44k spots (22k spots printed twice). Thus, rep1 and rep2 are technical replicates of the same biological sample (derived from one raw data file).

ORGANISM(S): Magnaporthe oryzae

SUBMITTER: Karsten Andresen

PROVIDER: E-GEOD-36224 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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The role of the Tra1p transcription factor of Magnaporthe oryzae in spore adhesion and pathogenic development

Project description:This SuperSeries is composed of the following subset Series: GSE36224: Comparison of transcript abundance in aerial mycelium of the Magnaporthe oryzae TRA1-deleted mutant and its parental strain GSE36225: Comparison of transcript abundance in ungerminated spores of the Magnaporthe oryzae TRA1-deleted mutant and its parental strain Refer to individual Series

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Project description:Promoter-specific transcriptional activators (activators) stimulate transcription through direct interactions with one or more components of the transcription machinery, termed the “target”. Previous studies have provided evidence that the Tra1 subunit of the yeast SAGA (Spt-Ada-Gcn5-acetyltransferase) complex is the target of the yeast activator Gal4. However, several other general transcription factors, in particular the mediator complex, have also been implicated as Gal4 targets. To investigate the essentiality of Tra1 as a target of Gal4, here we derive Tra1 mutants that are selectively defective for interaction with Gal4 in vivo (Gal4 Interaction Defective (GID) mutants). In contrast to wild-type Tra1, Tra1 GID mutants are not recruited by Gal4 to the promoter and cannot support Gal4-directed transcription activation, demonstrating that the Gal4–Tra1 interaction is required for Gal4 function. In yeast strains expressing a Tra1 GID mutant, Gal4 promoter binding is unexpectedly also diminished indicating that Gal4 and Tra1 bind cooperatively. Consistent with cooperative binding, we demonstrate that the interaction between Gal4 and Tra1 occurs predominantly on the promoter and not off DNA. Finally, we show that although Tra1 is also targeted by other activators, these interaction are unaffected by GID mutations, revealing an unanticipated specificity of the Gal4-Tra1 interaction.

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Gene profiling of TRA1 and tra1-L3733A

Project description:Tra1 is a component of the Saccharomyces cerevisiae SAGA and NuA4 complexes and a member of the phosphatidylinositol 3-kinase kinase family that contain a C-terminal PI3K domain followed by a 35-residue FATC domain. We analyzed alleles with single residue changes in the FATC domain. Of these tra1-L3733A had the most pronounced effects, resulting in phenotypes similar but not identical to those resulting from mutations in the PI3K domain or SAGA or NuA4 components. To analyze the transcriptional effects of the tra1-L3733A allele, we performed gene profiling of wild-type and tra1-L3733A strains. Expression of 11 genes was elevated ≥ 2-fold, 79 genes were decreased ≥ 2-fold. Of the 11 genes with elevated expression, four (HSP26, PIR3, DDR2 and GRE1) have roles in cellular stress response. The only gene ontology process significantly enriched in the genes with decreased expression greater than 2-fold was lysine biosynthesis (GO: 0009085). Broader analysis of gene ontology processes revealed expression of genes involved in cellular respiration (GO:0045333) and the overlapping process generation of precursor metabolites and energy (GO: 0006091) showed an average upward shift of ~10% relative to the total collection of genes. A similar trend of less magnitude was seen for vitamin metabolic processes (GO:0006766). A slight downward shift was seen for genes involved in translation (GO:0006412) and membrane organization (GO:0016044).

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