Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Studies of target genes for signal transduction pathways (using a Ras1-, a Cdc42- and a Gap1-mutant) in the basidiomycete Schizophyllum commune.

ABSTRACT: 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.

ORGANISM(S): Schizophyllum commune

SUBMITTER: Erika Kothe

PROVIDER: E-GEOD-43965 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

A central role for Ras1 in morphogenesis of the basidiomycete Schizophyllum commune.

Knabe Nicole N Jung Elke-Martina EM Freihorst Daniela D Hennicke Florian F Horton J Stephen JS Kothe Erika E

Eukaryotic cell 20130419 6

Fungi have been used as model systems to define general processes in eukaryotes, for example, the one gene-one enzyme hypothesis, as well as to study polar growth or pathogenesis. Here, we show a central role for the regulator protein Ras in a mushroom-forming, filamentous basidiomycete linking growth, pheromone signaling, sexual development, and meiosis to different signal transduction pathways. ras1 and Ras-specific gap1 mutants were generated and used to modify the intracellular activation st ...[more]

PMID: 23606288

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Project description:MicroRNAs (miRNAs) are a class of endogenous, non-coding RNAs that mediate post-transcriptional gene silencing by inhibiting mRNA translation and promoting mRNA decay. DICER1, an RNAse III endonuclease encoded by Dicer1, is required for processing short 21-22 nucleotide miRNAs from longer double-stranded RNA precursors. Here, we investigate the loss of Dicer1 in mouse postnatal male germ cells to determine how disruptions in the miRNA biogenesis pathway may contribute to infertility. Reduced levels of Dicer1 transcripts and DICER1 were confirmed in germ cell knock-out (GCKO) testes by postnatal day 18 (P18). Compared to wild-type (WT) at 8 weeks, GCKO males had no change in body weight, yet showed significant reductions in testis mass and sperm number. Histology and fertility tests confirmed spermatogenic failure in GCKO males. Array analyses at P18 showed 96% of miRNA genes were down-regulated and 37% of protein-coding genes were differentially expressed in GCKO testes. Interestingly, we observed preferential overexpression of genes on the sex chromosomes in GCKO testes, with more than 80% of the genes overlapping those proposed to undergo meiotic sex chromosome inactivation (MSCI) in the germ cells. Compared to WT, GCKO mice showed higher percentages of cells at early meiotic stages (leptotene and zygotene) but lower percentages at later stages (pachytene, diplotene and metaphase I), providing evidence that deletion of Dicer1 leads to disruptions in meiotic progression. Furthermore, we observed fewer elongating spermatids with proper translational activation of transition protein 2 (Tnp2), protamine 1 and 2 (Prm1 and Prm2) in GCKO testes after step 12-14. Therefore, deleting Dicer1 in early postnatal germ cells causes misregulation of transcripts encoded by genes on the sex chromosomes, impairs meiotic progression and post-meiotic translational control and results in spermatogenic failure and infertility. Total RNA, including miRNAs, were purified from a total of six individual mouse samples. The tissue collected was obtained from wild-type (control; n=3) and Dicer1 germ cell knockout (mutant; n=3) testes on P18. One miRNA GCKO sample, M36, was determined to be of poor quality and was excluded from the study; thus, a total of five miRNA samples were analyzed.

Dataset Information

Studies of target genes for signal transduction pathways (using a Ras1-, a Cdc42- and a Gap1-mutant) in the basidiomycete Schizophyllum commune.

Publications

A central role for Ras1 in morphogenesis of the basidiomycete Schizophyllum commune.

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