Dataset Information

Salmonella mediated intracellular niche control by SCV size regulation

ABSTRACT: Salmonella is a human and animal pathogen causing gastro-enteric diseases worldwide. The key feature of Salmonella infection is its entry into intestinal epithelial cells within a Salmonella-Containing Vacuole (SCV). This original compartment is distinct from empty macropinosomes formed around the infection site. A few minutes after its formation, the SCV increases in size through fusions with the surrounding macropinosomes. On the opposite, Salmonella induces the formation of elongated tubules leading to SCV membrane and volume loss. Later, the SCV can mature into a vacuolar pathogen niche, or be ruptured releasing Salmonella in the host cytosol where the bacteria hyper-replicates. Here, we describe how size control of the early SCV is the main contributor to its stability and consequently determines the Salmonella intracellular niche and growth. We identify the SNAREs required for increasing the SCV size through fusions. We show that this fusion promotes the maintenance of the SCV integrity and the establishment of a vacuolar niche

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Salmonella Enterica Subsp. Enterica Serovar Typhimurium

DISEASE(S): Gastroenteritis

SUBMITTER: Magalie Duchateau

LAB HEAD: Jost Enninga

PROVIDER: PXD012825 | Pride | 2020-01-09

REPOSITORIES: Pride

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Publications

Dynamic Growth and Shrinkage of the Salmonella-Containing Vacuole Determines the Intracellular Pathogen Niche.

Stévenin Virginie V Chang Yuen-Yan YY Le Toquin Yoann Y Duchateau Magalie M Gianetto Quentin Giai QG Luk Chak Hon CH Salles Audrey A Sohst Victoria V Matondo Mariette M Reiling Norbert N Enninga Jost J

Cell reports 20191201 12

Salmonella is a human and animal pathogen that causes gastro-enteric diseases. The key to Salmonella infection is its entry into intestinal epithelial cells, where the bacterium resides within a Salmonella-containing vacuole (SCV). Salmonella entry also induces the formation of empty macropinosomes, distinct from the SCV, in the vicinity of the entering bacteria. A few minutes after its formation, the SCV increases in size through fusions with the surrounding macropinosomes. Salmonella also indu ...[more]

PMID: 31851926

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Project description:Intracellular bacterial pathogens inject a cocktail of effector proteins into host cells to hijack diverse cellular processes and promote their survival and proliferation. Salmonella enterica serovar Typhimurium (STm) uses two Type 3 secretion system (T3SS) to deliver a suite of effector proteins into host cells, however, knowledge of their host-targets remains sparse. To systematically map STm effector-host protein-protein interactions (PPIs) during active infection, we generated a library of 32 STm strains expressing affinity-tagged effector proteins from their endogenous chromosomal loci. Tagged-effector strains were then used to infect macrophages or epithelial cells, and PPIs were detected by Affinity-Purification Quantitative Mass-Spectrometry (AP-QMS) under both native and cross-linking conditions. We recovered 25 previously described effector-host PPIs, along with 421 novel interactions across the two cell lines tested, as well as several effector-effector PPIs. While effectors converged on specific host cellular processes, which varied across cell types, most had multiple host targets. Using reciprocal co-immunoprecipitations, we could validated 13 out of 22 new PPIs, demonstrating an accuracy of at least 59% for the AP-QMS approach. To illustrate the utility of this resource for discovering novel infection biology, we first showed that SseJ and SseL cooperate to control cholesterol transport at the Salmonella Containing Vacuole (SCV) via the Niemann-Pick C1 protein (NPC1). Second, we uncovered that PipB directly binds and recruits the organelle contact site protein PDZD8 to the SCV. Third, we elucidated a novel mechanism whereby the effector kinase SteC promotes host actin bundling, by directly interacting and phosphorylating formin-like proteins. Overall, we provide a systems-wide host-bacterial pathogen physical interactome resource, to our knowledge the first in an infection context, with broad implications for effector function and cooperation.

Project description:Abstract Barley is a cereal crop of worldwide importance for brewing industry, animal and in lesser extent humannutrition. Grain development is a tightly controlled process which predominantly contributes to the final grain quality. To understand molecular changes occurring in the early seed development stage up to the initial storage phase, proteome profiling of developing barley seed at 8, 10 18 and 20 days after flowering was analyzed. In vivo label free quantification approach allowed the quantification of more than 1200 proteins across the different stages. Confirmation of the reproducibility of the developmental data obtained in our proteomics analysis was performed using western-blot technics to validate the specific pattern of selected proteins and comparison with previously published data. Multi- and uni-variates statistics were used to unravel the biochemical dynamics underlying the grain filling process. Seed storage proteins (SSPs) were found to accumulate during the grain filling process suggesting a higher activity of the endoplasmic reticulum playing a major role in protein synthesis. This hypothesis was supported by a spatio-temporal analysis of the ER marker protein-disulfide-isomerase (PDI) at both transcript and protein level coupled with microscopic study of the ER dynamics. Results indicates that ER is predominantly active in the starchy endosperm up to 18 DAP and is subsequently degraded while the cells undergo program cell death. The central role of the ER was further strengthening by the analysis of all ER related proteins. Similarly, the pattern of vacuolar H+ ATPases suggested a decrease of the vacuolar acidity, microscopic analysis of the endosperm unravel a concomitant decrease of both vacuolar size and acidity associated with an abundance switch in different class of vacuolar annotated proteins (eg: Sucrose related metabolism proteins). Observed endomembrane system dynamics were supported by specific abundance changes of ESCRT related proteins such as VPS4 and SNF7 or VPS5 a protein associated to the retromer complex. Collectively, our results provide insight into endomembrane molecular processes taking place during barley early grain development, additionally the proteome dataset provide a comprehensive overview of a wide range of metabolic pathway which will be helpful for the cereal community.

Dataset Information

Salmonella mediated intracellular niche control by SCV size regulation

Publications

Dynamic Growth and Shrinkage of the Salmonella-Containing Vacuole Determines the Intracellular Pathogen Niche.

Similar Datasets

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