Project description:This SuperSeries is composed of the following subset Series: GSE32542: Murine serum reactivity to common autoantigens in response to immunization with neutrophil extracellular traps GSE32543: Human and murine serum reactivity to specific histone posttranslational modifications in neutrophil extracellular traps Refer to individual Series
Project description:As a dynamic structure with a barrier between the cell and the outside world, maintenance of cell wall integrity (CWI) is important for fungal growth, development and pathogenicity processes. Here we characterized a MADS-box transcription factor RlmA (AoRlmA) downstream of the CWI regulatory pathway in the nematode-trapping fungus Arthrobotrys oligospora. Deletion of AorlmA caused a reduction in mycelial growth and the number of nuclei, and significant downregulation of transcript levels of genes related to nucleus synthesis and DNA damage repair, such as rad3, spo11, and rad25. Meanwhile, the ΔAorlmA mutant strains showed a significant reduction in spore production compared with the wild-type (WT) strain, and the transcript levels of sporulation-related genes was downregulated in the ΔAorlmA mutant during the early and middle stages of condiation, such as flbA, medA, and vosA. Meanwhile, the mycelial cell wall of the ΔAorlmA mutant strain showed breakage. Also, the transcript levels of genes related to cell wall synthesis were significantly down-regulated in the mutant strain compared to the WT strain, and the mutant strain was more sensitive to stresses of cell wall synthesis disruptors, oxidative stress and osmotic stress than the WT strain. Compared with the WT strain, the ΔAorlmA mutant strain not only had a reduced number of traps and nematicidal ability, but also, the shape of the traps of the mutants has also changed. In addition, In addition, AoRlmA also regulates autophagy and endocytosis. Transcriptome analysis of ΔAorlmA mutant strains showed that AorlmA regulates redox, cell wall synthesis, DNA replication and damage repair, and pathogenic processes. Metabolomic analysis showed that AorlmA is involved in the biosynthesis of secondary metabolites of A. oligospora. To summarise, our data suggest that AorlmA is involved in growth, sporulation, spore germination, maintenance of cell wall integrity, DNA replication and damage repair, pathogenesis, autophagy and secondary metabolite biosynthesis processes.
Project description:Spores of Bacillales and Clostridiales species contain 100s of different mRNAs, and their major function in Bacillus subtilis is to provide ribonucleotides for new RNA synthesis when spores germinate. In new work, RNA was isolated from spores of five Bacillales and one Clostridioides species and relative spore mRNA levels were determined by RNA-seq. Determination of RNA levels in single spores allowed calculation of RNA nt/spore, and assuming mRNA is 3% of spore RNA allowed calculation that only ~6% of spore mRNAs were present at ≥ 1/spore. Bacillus subtilis, Bacillus atrophaeus and Clostridioides difficile spores had 49, 42 and 51 mRNAs at >1/spore, respectively. Numbers of mRNAs at ≥1/spore were ~10 to 50% higher in Geobacillus stearothermophilus and Bacillus thuringiensis Al Hakam spores, respectively, and ~ 4-fold higher in Bacillus megaterium spores. Notably, in all species: i) many of the 60 most abundant spore mRNAs were transcribed by RNA polymerase with forespore-specific s factors; ii) some to many of the most abundant spore mRNAs encoded orthologs of those encoded by abundant B. subtilis spore mRNAs and proteins present in dormant spores ; and iii) some spore mRNAs were likely transcribed in the mother cell compartment of the sporulating cell. Indeed , analysis of the coverage of RNA-seq reads on mRNAs from all six species suggested that abundant spore mRNAs were at least somewhat fragmented. This observation was confirmed by RT-qPCR analysis of three abundant mRNAs each from B. subtilis and C. difficile spores. These data add to a growing body of evidence indicating that the great majority of mRNAs in spores of Firmicutes are degradation and function as a ribonucleotide depot for new RNA synthesis when spores germinate.
Project description:Bacillus subtilis forms dormant spores upon nutrient depletion. Under favorable environmental conditions, the spore breaks its dormancy and resumes growth in a process called spore germination and outgrowth. To elucidate the physiological processes that occur during the transition of the dormant spore to an actively growing vegetative cell, we studied this process in a time-dependent manner by a combination of microscopy, analysis of extracellular metabolites and a genome-wide analysis of transcription. The results indicate the presence of abundant levels of late sporulation transcripts in dormant spores. In addition, results suggest the existence of a complex and well-regulated spore outgrowth program, involving the temporal expression of at least 30 % of the B. subtilis genome. Keywords: time course, spore outgrowth
Project description:A microarray analysis was conducted to investigate transcriptional differences between traps and vegetative hyphae of the nematophagous fungus Monacrosporium haptotylum. Our goal was to identify genes that show differential regulation in the two different cell types; 1) traps (knobs) and 2) vegetative hyphae (mycelium) grown in the same medium. The entire design involved nine slides; four of these were hybridisations of knobs versus mycelium and five were mycelium against mycelium, 10 labelled extracts, and 3 biological replicates. The microarray experiments were designed as two-sample comparisons (i.e. knobs versus mycelium or mycelium versus mycelium) using three independent biological replicates, which also included technical and dye-swapped control hybridisations.
Project description:To gain insight into spore germination and outgrowth, the transcriptome changes during Bacillus subtilis spore conversion to vegetative cells were analyzed. The transcriptome analysis also allowed us to trace the different functional groups of genes expressed during this conversion. . Our analysis identified 34 abundant mRNA transcripts in the dormant spores, at least 31 of which were rapidly degraded after the phase transition and observed 3152 differentially expressed genes during spore germination and outgrowth.
Project description:spaA is a Cud-type transcription factor that is essential for spore cell differentiation of Dictyostelium discoideum, a social amoeba. ChIP-seq was performed to identify spaA target genes.
Project description:In filamentous fungi, asexual sporulation involves morphological differentiation and metabolic changes. The process of asexual spore formation is tightly regulated by a variety of transcription factors including VosA, VelB, and WetA. A number of studies have demonstrated that these three transcription factors are key regulators of asexual spore formation and maturation in the model filamentous fungus Aspergillus nidulans. To gain a more mechanistic view of the roles these transcription factors play in asexual spores, genome-wide and metabolomic analyses were conducted in A. nidulans conidia. RNA sequencing and chromatin immunoprecipitation-based sequencing data suggested that the three transcription factors directly or indirectly regulate the expression of genes associated with spore-wall integrity, asexual development, and secondary metabolism. In addition, metabolomics analysis of conidia extracts showed strikingly different primary and secondary metabolite profiles for wild-type and mutant conidia. These results suggest that WetA, VosA, and VelB play key roles in the morphological development of and metabolic changes in conidia. This entry is for the ChIP-seq data.