ABSTRACT: Studies of target genes for signal transduction pathways (using a Ras1-, a Cdc42- and a Gap1-mutant) in the basidiomycete Schizophyllum commune.
Project description:Intracellular signaling is conserved in eukaryotes to allow for response to extracellular signals and to regulate development and cellular functions. In fungi, inositol phosphate signaling has been shown to be involved in growth, sexual reproduction, and metabolic adaptation lacking, however, reports of mushroom forming fungi so far. In Schizophyllum commune, an inositol monophosphatase has been found up-regulated during sexual development. The enzyme is crucial for inositol cycling where it catalyzes the last step of inositol phosphate metabolism restoring the inositol pool from the monophosphorylated inositol monophosphate. We overexpressed the gene in this model basidiomycete and verified its involvement in cell wall integrity and intracellular trafficking. Strong phenotypes in mushroom formation and cell metabolism were evidenced by proteome analyses. In addition, altered inositol signaling was shown to be involved in tolerance towards cesium and zinc, and increased metal tolerance towards cadmium, associated with induced expression of kinases and repression of phosphatases within the inositol cycle. The presence of the heavy metals Sr, Cs, Cd, and Zn lowered intracellular calcium levels. We could develop a model integrating inositol signaling in the known signal transduction pathways governed by Ras, G-protein coupled receptors, cAMP and elucidate different roles in development.
Project description:Transcriptome changes associated with signal transduction mutants were analysed in order to identify targets of the distinct pathways of Ras1, Cdc42 and Gap1. Two monokaryons (12-43 and 4-39), a dikaryon (W22 x 12-43), a constitutive active Ras1 mutant (T2G12V, monokaryon), a Cdc42 mutant (G12V, monokaryon) and a M-bM-^HM-^Fgap1 x M-bM-^HM-^Fgap1 dikaryon (F15 x F28, deletion of Gap1) were analyzed for that purpose. With microarray analysis mRNA expression in different mutants of the basidiomycete S. commune was analyzed. Genes which show regulation in the comparison either of the two monokaryons 12-43 and 4-39 (for Ras2 and Cdc42 mutant) or the dikaryon W22 x 12-43 (for Gap1 x Gap1 dikaryon) have been eliminated from other comparisons as stage-specific (mono- or dikaryon) differences. Two biological replicate samples were analyzed for each condition.
Project description:The transcription factors Hom2 and Fst4 were shown to be involved in mushroom development in the basidiomycete Schizophyllum commune. When the genes encoding Hom2 or Fst4 were deleted, no mushrooms formed. In this study we performed a genome-wide expression analysis on dikaryons of wild-type, delta-hom2 and delta-fst4.
Project description:The transcription factors Hom2 and Fst4 were shown to be involved in mushroom development in the basidiomycete Schizophyllum commune. When the genes encoding Hom2 or Fst4 were deleted, no mushrooms formed. In this study we performed a genome-wide expression analysis on dikaryons of wild-type, delta-hom2 and delta-fst4. A total of three samples were analyzed. One wild-type and two deletion strains (delta-hom2 and delta-fst4)
Project description:In order to study the effect of light on gene expression in Schizophyllum commune, genome wide gene expression was analysed in 4-day-old monokaryotic and dikaryotic wild type colonies, grown either in light or in darkness.
Project description:In order to study the effect of light on gene expression in Schizophyllum commune, genome wide gene expression was analysed in 4-day-old monokaryotic and dikaryotic wild type colonies, grown either in light or in darkness. 4 samples: - monokaryon, grown for 4 days in the light - monokaryon, grown for 4 days in the dark - dikaryon, grown for 4 days in the light - dikaryon, grown for 4 days in the dark RNA was obtained from 3 biological replicates and pooled
