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Grosboillot, V.

Publications and source records attributed to Grosboillot, V..

2 recordsLinked to original sources

Competence repression by the SPβ prophage in Bacillus subtilis revealed by transcriptomics across culture conditions

Prophages are near-ubiquitous in bacterial genomes, yet their effects on host physiology are typically assayed under a narrow range of laboratory conditions, mostly involving DNA-damaging agents or physical and chemical stress. This leaves open the question of which ecological and physiological conditions trigger prophage-host interaction, and whether the outcome benefits the host or the prophage. Here, we used publicly available transcriptomic data to investigate conditions promoting prophage-host interaction in Bacillus subtilis. This analysis identified that competence strongly induces prophage gene expression. To further investigate the role of competence, we used two sets of B. subtilis strains: a natural SP{beta} lysogen strain and its cured counterpart, and a natural non-lysogenic strain and the same strain lysogenised with SP{beta}. Under competence-inducing conditions, both lysogen strains showed lower expression of the competence genes compared with their non-lysogenic counterparts. This result was corroborated by transformation assays that showed consistently lower transformation efficiency in the lysogen strains, and by fluorescence microscopy using a transcriptional reporter for the late competence gene. Because competence is a principal route for horizontal gene transfer in B. subtilis, prophage repression of the competence programme constrains host DNA uptake and evolvability. More broadly, these results show that mining public transcriptomes for condition effects can guide experimental effort towards prophage-host phenotypes that conventional laboratory conditions would miss.

microbiology↗

A proteogenomic resource enabling integrated analysis of Listeria genotype-proteotype-phenotype relationships

Listeria monocytogenes is an opportunistic foodborne pathogen responsible for listeriosis, a potentially fatal foodborne disease. Many different Listeria strains and serotypes exist, but a proteogenomic resource that bridges the gap in our molecular understanding of the relationships between the Listeria genotypes and phenotypes via proteotypes is still missing. Here we devised a next-generation proteogenomics strategy that enables the community to rapidly proteotype Listeria strains and relate this information back to the genotype. Based on sequencing and de novo assembly of the two most commonly used Listeria model strains, EGD-e and ScottA, we established two comprehensive Listeria proteogenomic databases. A genome comparison established core- and strain-specific genes potentially responsible for virulence differences. Next, we established a DIA/SWATH-based proteotyping strategy, including a new and robust sample preparation workflow, that enables the reproducible, sensitive, and relative quantitative measurement of Listeria proteotypes. This reusable and publically available DIA/SWATH library covers 70% of open reading frames of Listeria and represents the most extensive spectral library for Listeria proteotype analysis to date. We used these two new resources to investigate the Listeria proteotype in states mimicking the upper gastrointestinal passage. Exposure of Listeria to bile salts at 37 {degrees}C, which simulates conditions encountered in the duodenum, showed significant proteotype perturbations including an increase of FlaA, the structural protein of flagella. Given that Listeria is known to lose its flagella above 30 {degrees}C, this was an unexpected finding. The formation of flagella, which might have implications on infectivity, was validated by parallel reaction monitoring and light and scanning electron microscopy. flaA transcript levels were not significantly different with and without exposure to bile salts at 37 {degrees}C, suggesting regulation at the post-transcriptional level. Together, these analyses provide a comprehensive proteogenomic resource and toolbox for the Listeria community enabling the analysis of Listeria genotype-proteotype-phenotype relationships.

systems biology↗