Project description:To understand which gene(s) are affected by 2-methylcitrate to lead to the delayed sporulation in ∆prpD, we extracted the total RNA from BMB171 and ∆prpD at 12 h for whole genome transcription analysis by RNA sequencing (RNA-Seq). Our RNA-seq data showed that transcriptions of almost all of the SigF regulon genes, as concluded from the B. subtilis regulon, were significantly down-regulated in ∆prpD compared with the wild type BMB171. However, almost all the regulon genes of SigH, a sigma factor that functions upstream of SigF, were significantly up-regulated in ∆prpD. Those genes included spo0A, the early-expressed operon spoIIA and sigF itself. These results indicated that dysregulation of genes occurred on various levels. In particular, though, post-transcriptional inhibition of sigF functionality might subsequently lead to down-regulation of SigF regulon genes in the ∆prpD mutant.
Project description:Transcriptional changes during asexual sporangia formation by the late blight pathogen Phytophthora infestans were identified using microarrays representing 15,650 genes and RNA from sporulation time-courses, purified spores, and sporulation-defective strains. Results were confirmed by reverse transcription-polymerase chain reaction analyses of sporulation on artificial media and infected tomato. During sporulation, about 12% of genes were found to be up-regulated and 5% down-regulated. The most prevalent induced genes had functions in signal transduction, flagella assembly, cellular organization, metabolism, and molecular or vesicular transport. Distinct patterns of expression were discerned based on the kinetics of mRNA induction and their persistence in sporangia. For example, most flagella-associated transcripts were induced very early in sporulation and maintained in sporangia, while many participants in metabolism or small molecule transport were also induced early but had low levels in sporangia. Data from this study are a resource for understanding sporogenesis, which is critical to the pathogenic success of P. infestans and other oomycetes.
Project description:Endospore (hereafter called spore) is a dormant, tough, and non-reproductive structure produced in a lack of nutrients by certain species of bacteria from the Firmicute phylum. Sporulation is tightly linked to cell cycle and involving by temporal and spatial regulation of hundreds genes. It has been suggested more than 500 genes are contributed to sporulation in B. subtilis, and there are certainly more. In our study, we identified four new sporulation genes and their mutants exhibited sporulation abnormal. RNA-seq of mutants compared to wild type during sporulation will reveal the regulation of sporulation related genes in the four mutants, and therefore shed the light to the functional role of the four new sporulation genes.
Project description:The aim of this study was to expand our knowledge about the gene repertoire involved in S. coelicolor sporulation. Here we compare the global transcription patterns of the wildtype (M145) parent and two mutants lacking key regulators of aerial hyphal sporulation (whiA and whiH).
Project description:Blastocladiella emersonii life cycle presents a number of drastic biochemical and morphological changes, mainly during two cell differentiation stages: the germination and the sporulation. To investigate the transcriptional changes taking place during the sporulation phase, which culminates with the production of the zoospores, motile cells responsible for the dispersal of the fungus, microarray experiments were performed.
Project description:Translational control during the intricate process of sporulation in Bacillus subtilis as a response to nutrient limitation is still underexplored. Here, we employed a comprehensive approach including RNA-seq, ribosome profiling and fluorescence microscopy to dissect the translational landscape of B. subtilis during sporulation. We identified two events of translation silencing and described the spatiotemporal changes in the subcellular location of translational machinery during sporulation. Using a triple knock-out strain (3KO) of zinc-independents paralogs of three zinc-dependent ribosomal proteins L31, L33 and S14, we investigated the potential regulatory role of ribosome during sporulation. The 3KO strain exhibited delayed sporulation, reduced germination efficiency, and dysregulated translation including expression of key metabolic and sporulation-related genes as well as disruptions in translation silencing, particularly in late sporulation.
Project description:Translational control during the intricate process of sporulation in Bacillus subtilis as a response to nutrient limitation is still underexplored. Here, we employed a comprehensive approach including RNA-seq, ribosome profiling and fluorescence microscopy to dissect the translational landscape of B. subtilis during sporulation. We identified two events of translation silencing and described the spatiotemporal changes in the subcellular location of translational machinery during sporulation. Using a triple knock-out strain (3KO) of zinc-independents paralogs of three zinc-dependent ribosomal proteins L31, L33 and S14, we investigated the potential regulatory role of ribosome during sporulation. The 3KO strain exhibited delayed sporulation, reduced germination efficiency, and dysregulated translation including expression of key metabolic and sporulation-related genes as well as disruptions in translation silencing, particularly in late sporulation.
Project description:The universally conserved protein Elongation Factor P facilitates translation at amino acids that form peptide bonds with low efficiency, particularly poly-proline tracts. Despite its wide conservation, it is not essential in most bacteria and its physiological role remains unclear. Here, we show that EF-P affects the process of sporulation initiation in the bacterium Bacillus subtilis. We observe that lack of EF-P delays expression of sporulation-specific genes. Using ribosome profiling, we observe that expression of spo0A, encoding a transcription factor that functions as the master regulator of sporulation, is lower in a ∆efp strain as compared to the wildtype. Ectopic expression of Spo0A rescues the sporulation initiation phenotype, indicating that reduced spo0A expression explains the sporulation defect in ∆efp cells. Since Spo0A is the earliest sporulation transcription factor, these data suggest that sporulation initiation can be delayed when protein synthesis is impaired.
Project description:In Bacillus subtilis, sporulation is a sequential and highly regulated process. Phosphorylation events by Histidine or Serine/Threonine kinases are key points in this regulation. PrkA has been proposed to be an essential Serine kinase for the initiation of sporulation but its kinase activity has not been clearly demonstrated so far. Indeed, neither its autophosphorylation nor identification of a B. subtilis phosphorylated substrate was unambiguously established. Bioinformatic homology searches revealed sequence similarities with the AAA+ ATP-dependent Lon protease family. Here, we showed that PrkA is indeed able to hydrolyse the α-casein, an exogenous substrate of Lon proteases, in an ATP-dependent manner. We also showed that this ATP-dependent protease activity is essential for PrkA function in sporulation since mutation in the Walker A motif leads to a sporulation defect. Furthermore, we found that PrkA protease activity is tightly regulated by phosphorylation events involving one of the Ser/Thr kinases of B. subtilis, PrkC, as characterized by mass spectrometry from 3 in vitro independent experiments (1: PrkA+PrkC/2:PrKA-S219E+PrkC/3: PrKA-T217E+PrkC). We finally demonstrated that PrkA regulation of the transcriptional factor σK via the transition phase regulator ScoC is certainly indirect.