Dataset Information

Salmonella temperature dependent proteome

ABSTRACT: Salmonella enterica serovar Typhimurium (STM) is one of the major causes of gastroenteritis and is linked to the consumption of contaminated food. Thereby, STM is associated to food of animal related produce (pork, chicken, eggs) and to food of non-animal related produce (vegetables, fruits, nuts, herbs and water). A lot of studies investigated the process of infection of warm-blooded mammalian hosts by STM and the underlying complex regulatory network of virulence gene expression dependent of various environmental conditions encountered in hosts. However, less is known about the proteome and possible regulatory networks for gene expression of STM outside the preferred host. Nutritional limitations and changes in temperature are the most obvious stresses outside the native host. Thus, we analyzed the proteome profile of STM grown in rich medium (LB medium) and minimal medium (PCN medium) at temperatures ranging from 8 °C to 37 °C. LB medium mimics the nutritional rich environment inside the host, whereas minimal PCN medium represents nutritional limitations outside the host, found during growth of non-animal related produce (field conditions). Further, the used range of temperature is found inside natural hosts (37 °C), at ambient room conditions (20 °C), during agricultural growth on open field (16 °C and 12 °C) and probably during food storage (8 °C). For the investigation of implications of altered nutrient availability and changes in growth temperature on STM proteome, whole cell lysates of STM grown under various conditions were analyzed by HPLC/MS-MS and label-free quantification. With this method, we are able to decipher changes in the complete proteome of STM in comparison to standard laboratory conditions mimicking the natural host.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Salmonella Typhimurium (strain 14028s / Sgsc 2262)

DISEASE(S): Salmonellosis

SUBMITTER: Joerg Deiwick

LAB HEAD: Michael Hensel

PROVIDER: PXD025848 | Pride | 2022-07-26

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	STM14028_5.fasta	Fasta
	WT12LBWC-1-Wdh_190905_20190906105636.mgf	Mgf
	WT12LBWC-1-Wdh_190905_20190906105636.raw	Raw
	WT12LBWC-1-Wdh_190905_20190906105636_peptides_1_1_0.mzid.gz	Mzid
	WT12LBWC-2-Wdh_190905.mgf	Mgf

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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:As a broad group, hemibiotrophic pathogens cause significant losses within agriculture threatening the sustainability of food systems globally. The complex manner in which these microbes colonize their hosts, including an initial biotrophic phase of colonization followed by a biotrophic-to-necrotrophic switch (BNS) phase and ending with a necrotrophic mode of nutritional acquisition, renders their management more complex than pathogens which display a single nutritional strategy. Within model plant systems, typically each of these phases is characterized by both common and discrete transcriptional responses as the host modulates physiological processes in concert with changes in pathogen biology. Plant hormones also play an important role in these stages with classic models showing that salicylic acid accumulates during the biotrophic phase and jasmonic acid/ethylene responses occur during the necrotrophic phase. In this study, we use the interaction between the hemibiotroph Phytophthora medicaginis and the roots of its host Cicer arietinum (chickpea) to define the duration of the different life stages of P. medicaginis during the pathogenesis of chickpea. Using transcriptional profiling across all three stages of infection we demonstrate that chickpea displayed some similarities in response to P. medicaginis as has been previously documented in other model plant-pathogen hemibiotrophic interactions. However, our transcriptomic results suggest that chickpea does not conform to the phytohormone response model observed in leaf colonization, nor to that observed for roots of other plant species being colonised by soil-borne hemibiotrophs. We confirm these findings using targeted quantification of salicylic acid and jasmonic acid. The findings from this study demonstrate that a wider spectrum of plant species should be investigated in future to understand the physiological changes in plants during the different stages of colonization by hemibiotrophic soil-borne pathogens before we can better manage these economically important microbes in agronomically relevant conditions.

Project description:This model is from the article: Genetic and metabolic determinants of nutritional phenotype in an insect-bacterial symbiosis. MacDonald SJ, Thomas GH, Douglas AE. Mol Ecol. 2011 May;20(10):2073-84. 21392141 , Abstract: The pervasive influence of resident microorganisms on the phenotype of their hosts is exemplified by the intracellular bacterium Buchnera aphidicola, which provides its aphid partner with essential amino acids (EAAs). We investigated variation in the dietary requirement for EAAs among four pea aphid (Acyrthosiphon pisum) clones. Buchnera-derived nitrogen contributed to the synthesis of all EAAs for which aphid clones required a dietary supply, and to none of the EAAs for which all four clones had no dietary requirement, suggesting that low total dietary nitrogen may select for reduced synthesis of certain EAAs in some aphid clones. The sequenced Buchnera genomes showed that the EAA nutritional phenotype (i.e. the profile of dietary EAAs required by the aphid) cannot be attributed to sequence variation of Buchnera genes coding EAA biosynthetic enzymes. Metabolic modelling by flux balance analysis demonstrated that EAA output from Buchnera can be determined precisely by the flux of host metabolic precursors to Buchnera. Specifically, the four EAA nutritional phenotypes could be reproduced by metabolic models with unique profiles of host inputs, dominated by variation in supply of aspartate, homocysteine and glutamate. This suggests that the nutritional phenotype of the symbiosis is determined principally by host metabolism and transporter genes that regulate nutrient supply to Buchnera. Intraspecific variation in the nutritional phenotype of symbioses is expected to mediate partitioning of plant resources among aphid genotypes, potentially promoting the genetic subdivision of aphid populations. In this way, microbial symbioses may play an important role in the evolutionary diversification of phytophagous insects. This model originates from BioModels Database: A Database of Annotated Published Models (http://www.ebi.ac.uk/biomodels/). It is copyright (c) 2005-2011 The BioModels.net Team. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information. In summary, you are entitled to use this encoded model in absolutely any manner you deem suitable, verbatim, or with modification, alone or embedded it in a larger context, redistribute it, commercially or not, in a restricted way or not.. To cite BioModels Database, please use: Li C, Donizelli M, Rodriguez N, Dharuri H, Endler L, Chelliah V, Li L, He E, Henry A, Stefan MI, Snoep JL, Hucka M, Le Novère N, Laibe C (2010) BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. BMC Syst Biol., 4:92.

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Salmonella temperature dependent proteome

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