Project description:A whole genome microarray approach has been engaged in the heterothallic euascomycete Podospora anserina to identify genes that are differentially expressed beetwen a wild type kinetics of sexual reproduction and two mutant strains ∆RID and ∆SMR1.
Project description:Mutations in the gene for the mitochondrial matrix protease CLPP can cause human Perrault syndrome, which is characterized by male and female infertility, progressive sensorineural deafness, ataxia and leukoencephalopathy. This gene encodes a peptidase that is conserved since bacteria and localizes to mitochondrial matrix in eukaryotes. To compare and validate mouse and human findings, the filamentous fungus Podospora anserina was cultured and protein extracts were analyzed.
Project description:Aging of biological systems is controlled by various processes which have a potential impact on gene expression. Here we report a genome-wide transcriptome analysis of the fungal aging model Podospora anserina. Total RNA of three individuals of defined age were pooled and analyzed by SuperSAGE (serial analysis of gene expression). A bioinformatics analysis identified different molecular pathways to be affected during aging. While the abundance of transcripts linked to ribosomes and to the proteasome quality control system were found to decrease during aging, those associated with autophagy increase, suggesting that autophagy may act as a compensatory quality control pathway. Transcript profiles associated with the energy metabolism including mitochondrial functions were identified to fluctuate during aging. Comparison of wild-type transcripts, which are continuously down-regulated during aging, with those down-regulated in the long-lived, copper-uptake mutant grisea, validated the relevance of age-related changes in cellular copper metabolism. Overall, we (i) present a unique age-related data set of a longitudinal study of the experimental aging model P. anserina which represents a reference resource for future investigations in a variety of organisms, (ii) suggest autophagy to be a key quality control pathway that becomes active once other pathways fail, and (iii) present testable predictions for subsequent experimental investigations.
Project description:Here we report data of an analysis to monitor the age-related quantitative protein composition of the mitochondria of the fungal aging model Podospora anserina. The impact of senescence on mitochondrial protein composition was analyzed by LC-MS analysis. In an untargeted proteomic approach, we identified 795 proteins in samples from juvenile and senescent wild-type cultures and obtained quantitative information for 226 of these proteins by spectral counting. Despite the broad coverage of the proteome, no substantial changes linked to the aging process could be observed. LC-MS/MS-data were analyzed using the Sequest algorithm implemented in the Bioworks 3.3.1 software (Thermo scientific) for peptide identification versus a P. anserina protein database. As enzyme specificity Trypsin was selected. Two missed cleavages were permitted and the mass tolerance was set to 10 ppm. False discovery rates were estimated by the number of spectral matches to a decoy database. Acceptance criteria and filters were set to achieve a false positive rate of 5 %.