E-GEOD-51360 - A genetic and functional investigation of the Zn2Cys6 transcription factors RSE2 and RSE3 in Podospora anserina

Status
Released on 22 October 2013, last updated on 3 May 2014
Organism
Podospora anserina
Samples (56)
Array (1)
Protocols (6)
Description
Abstract In Podospora anserina, the two zinc cluster proteins RSE2 and RSE3 are essential for the expression of the gene encoding the alternative oxidase (aox) when the mitochondrial electron transport chain is impaired. In parallel they activated the expression of gluconeogenic genes encoding phosphoenolpyruvate carboxykinase (pck) and fructose-1,6-biphosphatase (fbp). Orthologues of these transcription factors are present in a wide range of filamentous fungi and no other role than the regulation of these three genes has been evidenced so far. In order to better understand the function and the organisation of RSE2 and RSE3, we conducted a saturated genetic screen based on the constitutive expression of the aox gene. We identified 10 independent mutations in 9 positions in rse2 and 11 in 5 positions in rse3. Deletions were generated at some of these positions and the effects analyzed. This analysis suggests the presence of central regulatory domains and a C-ter activation domain in both proteins. Microarray analysis revealed 598 genes that were differentially expressed in the strains containing gain- or loss-of-function mutations in rse2 or rse3. It showed that in addition of aox, fbp and pck, RSE2 and RSE3 regulate the expression of genes encoding the alternative NADH dehydrogenase, a Zn2Cys6 transcription factor, a flavohemoglobin and various hydrolases. As a complement to expression data, a metabolome profiling approach revealed that both an rse2 gain-of-function mutation and growth on antimycin result in similar metabolic alterations in amino acids, fatty acids and α-ketoglutarate pools. With GPL10116: 7 conditions each with four biological replicates: wt, rse2Y326D, rse3G642V, rse2Y326D-rse3G642V, Δrse2, Δrse3 and Δrse2-Δrse3; common reference is a pool of four conditions M48h, M96h, C48h and C96h ; Conditions are labelled in Cy3 and the common reference in Cy5
Experiment type
transcription profiling by array 
Contacts
Annie Sainsard-Chanet <annie.sainsard@cgm.cnrs-gif.fr>, Adeline Humbert, Carole H Sellem, Elodie Bovier
MIAME
PlatformsProtocolsFactorsProcessedRaw
Files
Investigation descriptionE-GEOD-51360.idf.txt
Sample and data relationshipE-GEOD-51360.sdrf.txt
Processed data (1)E-GEOD-51360.processed.1.zip
Array designA-GEOD-10116.adf.txt
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