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Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation [RNA-seq]

ABSTRACT: 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.

ORGANISM(S): Bacillus subtilis

PROVIDER: GSE249449 | GEO | 2023/12/06

REPOSITORIES: GEO

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Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation [Ribo-seq]

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.

2023-12-06 | GSE249448 | GEO

Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation

Project description:The transcriptional control of sporulation in Bacillus subtilis is reasonably well understood, but its translational control is underexplored. Here, we use RNA-seq, ribosome profiling and fluorescence microscopy to study the translational dynamics of B. subtilis sporulation. We identify two events of translation silencing and describe spatiotemporal changes in subcellular localization of ribosomes during sporulation. We investigate the potential regulatory role of ribosomes during sporulation using a strain lacking zinc-independent paralogs of three zinc-dependent ribosomal proteins (L31, L33 and S14). The mutant strain exhibits delayed sporulation, reduced germination efficiency, dysregulated translation of metabolic and sporulation-related genes, and disruptions in translation silencing, particularly in late sporulation. This SuperSeries is composed of the SubSeries listed below.

2023-12-06 | GSE249450 | GEO

Elongation Factor P is important for sporulation initiation

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.

2023-01-04 | GSE222121 | GEO

Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation

Project description:Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation

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Sporulation in Bacillus subtilis (mother cell)

Project description:This series of experiments was designed to identify the program of gene transcription for a single differentiating cell type during sporulation in Bacillus subtilis. The mother cell is one of two cell types generated by asymmetric division of sporulating cells approximately two hours after initiation of sporulation. The program is governed by a hierarchical cascade consisting of the transcription factors: sigmaE, sigmaK, GerE, GerR (YlbO) and SpoIIID. The characterization of the sigmaE regulon was reported in Eichenberger et al. (2003), J. Mol. Biol. 327, 945-972. Here we report the data for sigmaK, GerE, GerR and SpoIIID.

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RNA-seq of four Bacillus subtilis mutants during sporulation

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.

2020-03-18 | E-MTAB-8124 | biostudies-arrayexpress

Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation [Ribo-seq]

Project description:Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation [Ribo-seq]

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Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation [RNA-seq]

Project description:Translation in Bacillus subtilis is spatially and temporally coordinated during sporulation [RNA-seq]

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High resolution time-lapse proteomic analysis of sporulation

Project description:Bacillus subtilis vegetative cells switch to sporulation upon nutrient limitation. To investigate the proteome changeover during sporulation, a time-lapse proteomic analysis was performed in a cell population that was induced to sporulate synchronously. Here, we are the first to comprehensively investigate the changeover of sporulation regulatory proteins, coat proteins and other proteins involved in sporulation and spore biogenesis. Protein co-expression analysis revealed four co-expressed modules. Modules brown and green are upregulated during sporulation and are annotated as associated with sporulation. Module blue, which is negatively correlated with modules brown and green, comprised ribosomal and metabolic proteins, and module yellow is co-expressed with module blue and modules brown & green. Notably, several proteins not belonging to any of the known transcription regulons were identified as co-expressed with modules brown and green. We speculate that they may also play roles during sporulation. Finally, amounts of some coat proteins, for example morphogenetic coat proteins, decreased late in sporulation. While we speculate on their possible role in guiding or helping assembly of other coat proteins, after which they can be disposed of, such a hypothesis remains to be experimentally addressed.

2021-04-03 | MSV000087154 | MassIVE

Global analysis of genes induced during sporulation of Clostridium difficile

Project description:Purpose: The goal of this study was to identify genes whose expression is induced during sporulation in a Spo0A-, σF-, σE-, σG-, and σK-dependent manner. Methods: Whole genome RNA sequencing was performed on wildtype, spo0A-, sigF-, sigE-, sigG-, and sigK- C. difficile strains (strain 630 background; JIR8094 = parent strain), and the transcriptional profiles of the different mutants during growth on 70:30 agar plates were determined using an Illumina HiSeq1000. Results: This analysis identified 185 genes whose expression is collectively activated by sporulation sigma factors: 150 were σF-dependent, 150 were σE-dependent, 30 were σG-dependent, and 31 were σK-dependent. A total of 237 genes were identified as requiring Spo0A for their expression when sporulation was induced on the 70:30 plates. Conclusions: These results provide the first genome-wide transcriptional analysis of genes whose expression is induced by specific sporulation sigma factors in the Clostridia and highlight that diverse mechanisms regulate sporulation sigma factor activity in the Firmicutes. For example, in contrast with the B. subtilis sporulation pathway, C. difficile σE was not required to fully activate σG, and σG was not required to activate σK.

2013-08-10 | GSE45977 | GEO

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