Transcriptional frameshifting rescues Citrobacter rodentium Type VI secretion by the production of two length variants from the prematurely interrupted tssM gene

The Type VI secretion system (T6SS) mediates toxin delivery into both eukaryotic and prokaryotic cells. It is composed of a cytoplasmic structure resembling the tail of contractile bacteriophages anchored to the cell envelope through a membrane complex composed of the TssL and TssM inner membrane proteins and of the TssJ outer membrane lipoprotein. The C-terminal domain of TssM is required for its interaction with TssJ, and for the function of the T6SS. In Citrobacter rodentium, the tssM1 gene does not encode the C-terminal domain. However, the stop codon is preceded by a run of 11 consecutive adenosines. In this study, we demonstrate that this poly-A tract is a transcriptional slippery site that induces the incorporation of additional adenosines, leading to frameshifting, and hence the production of two TssM1 variants, including a full-length canonical protein. We show that both forms of TssM1, and the ratio between these two forms, are required for the function of the T6SS in C. rodentium. Finally, we demonstrate that the tssM gene associated with the Yersinia pseudotuberculosis T6SS-3 gene cluster is also subjected to transcriptional frameshifting

Promoter swapping unveils the role of the Citrobacter rodentium CTS1 type VI secretion system in interbacterial competition.

International audienceThe type VI secretion system (T6SS) is a versatile secretion machine dedicated to various functions in Gram-negative bacteria, including virulence toward eukaryotic cells and antibacterial activity. Activity of T6SS might be followed in vitro by the release of two proteins, Hcp and VgrG, in the culture supernatant. Citrobacter rodentium, a rodent pathogen, harbors two T6SS gene clusters, cts1 and cts2. Reporter fusion and Hcp release assays suggested that the CTS1 T6SS was not produced or not active. The cts1 locus is composed of two divergent operons. We therefore developed a new vector allowing us to swap the two divergent endogenous promoters by P(tac) and P(BAD) using the λ red recombination technology. Artificial induction of both promoters demonstrated that the CTS1 T6SS is functional as shown by the Hcp release assay and confers on C. rodentium a growth advantage in antibacterial competition experiments with Escherichia coli

Étude de la thermosensibilité de transposition de la séquence d'insertion bactérienne IS911

TOULOUSE3-BU Sciences (315552104) / SudocSudocFranceF

The southern Tyrrhenian Sea margin: an example of lithospheric scale strike-slip duplex

The southern Tyrrhenian Sea margin is dominated by deformations whose kinematics are relatively poorly constrained, and different models have been proposed to account for its recent evolution. Analysis of new structural and space geodesy data, combined with available geophysical information, reveal a deformation field characterised by subhorizontal NW-SE directed shortening and SW-NE directed extension. The main recognised regional-scale structure comprises E-W trending fault zones, namely the Ustica-Eolie Line and the Mt. Kumeta-Alcantara Line, connected by the NW-SE trending Marettimo, Trapani, San Vito, Palermo, Gratteri-Mt. Mufara and Eolie faults. This fault network displays a remarkable dextral strike-slip character, and has the features of a strike-slip duplex, that we indicate as the Southern Tyrrhenian Strike-Slip Duplex (STSSD). Analysis of available geophysical data and regional considerations based on platetectonic reconstructions suggest that the STSSD is a primary feature of lithospheric-scale importance. Our strike-slip duplex model differs significantly from previously proposed passive margin interpretations for the Tyrrhenian Sea margin offshore northern Sicily. The results of this investigation show the effectiveness of an integrated, multidisciplinary approach in the study of active deformation margin

In vitro exploration of the Xanthomonas hortorum pv. vitians genome using transposon insertion sequencing and comparative genomics to discriminate between core and contextual essential genes

International audienceThe essential genome of a bacterium encompasses core genes associated with basic cellular processes and conditionally essential genes dependent upon environmental conditions or the genetic context. Comprehensive knowledge of those gene sets allows for a better understanding of fundamental bacterial biology and offers new perspectives for antimicrobial drug research against detrimental bacteria such as pathogens. We investigated the essential genome of Xanthomonas hortorum pv. vitians, a gammaproteobacterial plant pathogen of lettuce (Lactuca sativa L.) which belongs to the plant-pathogen reservoir genus Xanthomonas and is affiliated to the family Xanthomonadaceae. No practical means of disease control or preventionagainst this pathogen is currently available, and its molecular biology is virtually unknown. To reach a comprehensive overview of the essential genome of X. hortorum pv. vitians LM16734, we developed a mixed approach combining high-quality full genome sequencing, saturated transposon insertion sequencing (Tn-Seq) in optimal growth conditions, and coupled computational analyses such as comparative genomics, synteny assessment and phylogenomics. Among the 370 essential loci identified by Tn-Seq, a majority was bound to critical cell processes conserved across bacteria. The remaining genes were either related to specific ecological features of Xanthomonas or Xanthomonadaceae species, or acquired through horizontal genetransfer of mobile genetic elements and associated with ancestral parasitic gene behaviour and bacterial defence systems. Our study sheds new light on our usual concepts about gene essentiality and is pioneering in the molecular and genomic study of X. hortorum pv. vitians

A Comprehensive Overview of the Genes and Functions Required for Lettuce Infection by the Hemibiotrophic Phytopathogen Xanthomonas hortorum pv. vitians

International audienceThe successful infection of a host plant by a phytopathogenic bacterium depends on a finely tuned molecular cross talk between the two partners. Thanks to transposon insertion sequencing techniques (Tn-seq), whole genomes can now be assessed to determine which genes are important for the fitness of several plant-associated bacteria in planta. Despite its agricultural relevance, the dynamic molecular interaction established between the foliar hemibiotrophic phytopathogen Xanthomonas hortorum pv. vitians and its host, lettuce (Lactuca sativa), remains completely unknown. To decipher the genes and functions mobilized by the pathogen throughout the infection process, we conducted a Tn-seq experiment in lettuce leaves to mimic the selective pressure occurring during natural infection. This genome-wide screening identified 170 genes whose disruption caused serious fitness defects in lettuce. A thorough examination of these genes using comparative genomics and gene set enrichment analyses highlighted that several functions and pathways were highly critical for the pathogen's survival. Numerous genes involved in amino acid, nucleic acid, and exopolysaccharide biosynthesis were critical. The xps type II secretion system operon, a few TonB-dependent transporters involved in carbohydrate or siderophore scavenging, and multiple genes of the carbohydrate catabolism pathways were also critical, emphasizing the importance of nutrition systems in a nutrient-limited environment. Finally, several genes implied in camouflage from the plant immune system and resistance to immunity-induced oxidative stress were strongly involved in host colonization. As a whole, these results highlight some of the central metabolic pathways and cellular functions critical for Xanthomonas host adaptation and pathogenesis.IMPORTANCE Xanthomonas hortorum was recently the subject of renewed interest, as several studies highlighted that its members were responsible for diseases in a wide range of plant species, including crops of agricultural relevance (e.g., tomato and carrot). Among X. hortorum variants, X. hortorum pv. vitians is a reemerging foliar hemibiotrophic phytopathogen responsible for severe outbreaks of bacterial leaf spot of lettuce all around the world. Despite recent findings, sustainable and practical means of disease control remain to be developed. Understanding the host-pathogen interaction from a molecular perspective is crucial to support these efforts. The genes and functions mobilized by X. hortorum pv. vitians during its interaction with lettuce had never been investigated. Our study sheds light on these processes by screening the whole pathogen genome for genes critical for its fitness during the infection process, using transposon insertion sequencing and comparative genomics

The southern Tyrrhenian Sea margin: an example of lithospheric scale strike-slip duplex

The southern Tyrrhenian Sea margin is dominated by deforma- tions whose kinematics are relatively poorly constrained, and different models have been proposed to account for its recent evolution. Analy- sis of new structural and space geodesy data, combined with available geophysical information, reveal a deformation field characterised by subhorizontal NW-SE directed shortening and SW-NE directed exten- sion. The main recognised regional-scale structure comprises E-W trending fault zones, namely the Ustica-Eolie Line and the Mt. Kumeta-Alcantara Line, connected by the NW-SE trending Maret- timo, Trapani, San Vito, Palermo, Gratteri-Mt. Mufara and Eolie faults. This fault network displays a remarkable dextral strike-slip character, and has the features of a strike-slip duplex, that we indicate as the Southern Tyrrhenian Strike-Slip Duplex (STSSD). Analysis of available geophysical data and regional considerations based on plate- tectonic reconstructions suggest that the STSSD is a primary feature of lithospheric-scale importance. Our strike-slip duplex model differs significantly from previously proposed passive margin interpretations for the Tyrrhenian Sea margin offshore northern Sicily. The results of this investigation show the effectiveness of an integrated, multidisci- plinary approach in the study of active deformation margins