Dataset Information

Quantitative analysis of differential proteome expression in wild-type and mutants strains of Salmonella enterica serovar Typhimurium

ABSTRACT: The ability to adapt quickly to changing environmental conditions is crucial for growth and survival of bacteria in their natural environment. The common strategy that bacteria utilize to increase their survival under stressful conditions, including antibiotic treatment, is the entry into a non-actively growing state (quiescence). In the wide host-range pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium) and other Gram negative bacteria, this temporary arrest of proliferation induces the expression of the alternative sigma subunit σS/RpoS of RNA polymerase (RNAP) which remodels global gene expression to reshape the cell physiology and ensure survival in various stress conditions and nutritional deficiencies (i.e. general stress resistance) and long-term cell persistence. σS is also required for virulence and biofilm formation. Our work aims at understanding key molecular and physiological determinants responsible for persistence of non-growing bacteria, using the ubiquitous pathogen S. Typhimurium as a working system. We expect to identify novel functional proteins that are both physiologically important and of wide biological significance. One approach to characterize those proteins is to compare mutant proteome to that of the wild-type strain to unravel potential effects of the mutations on the expression and/or stability of proteins.

INSTRUMENT(S): Q Exactive HF

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

SUBMITTER: Thibaut Douché

LAB HEAD: Françoise Norel

PROVIDER: PXD005256 | Pride | 2018-10-26

REPOSITORIES: Pride

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			Action	DRS
	20160218_QHF_TD_ColES50_Norel_A1.raw	Raw
	20160218_QHF_TD_ColES50_Norel_A2.raw	Raw
	20160218_QHF_TD_ColES50_Norel_A3_160222152540.raw	Raw
	20160218_QHF_TD_ColES50_Norel_B1_160222215717.raw	Raw
	20160218_QHF_TD_ColES50_Norel_B2.raw	Raw

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Publications

Proteome remodelling by the stress sigma factor RpoS/σS in Salmonella: identification of small proteins and evidence for post-transcriptional regulation.

Lago Magali M Monteil Véronique V Douche Thibaut T Guglielmini Julien J Criscuolo Alexis A Maufrais Corinne C Matondo Mariette M Norel Françoise F

Scientific reports 20170518 1

The RpoS/σS sigma subunit of RNA polymerase is the master regulator of the general stress response in many Gram-negative bacteria. Extensive studies have been conducted on σS-regulated gene expression at the transcriptional level. In contrast, very limited information regarding the impact of σS on global protein production is available. In this study, we used a mass spectrometry-based proteomics approach to explore the wide σS-dependent proteome of the ...[more]

PMID: 28522802

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Project description:In many Gram-negative bacteria, the stress sigma factor of RNA polymerase, σS/RpoS, remodels global gene expression to reshape the physiology of quiescent cells and ensure their survival under non-optimal growth conditions. In the foodborne pathogen Salmonella enterica serovar Typhimurium, σS is also required for biofilm formation and virulence. We have recently shown that a ΔrpoS mutation affects the magnesium content of Salmonella and expression of the primary Mg2+-transporter CorA. The other two Mg2+-transporters of Salmonella are encoded by the PhoP-activated mgtA and mgtB genes expressed under magnesium starvation, whereas the corA gene is expressed constitutively. The σS control of corA prompted us to evaluate the impact CorA in quiescent Salmonella cells, by using global and analytical proteomic analyses and physiological assays. The ΔcorA mutation conferred a competitive disadvantage to exit from stationary phase, and slightly impaired motility, but had no effect on total and free cellular magnesium contents. In contrast to the wild type strain, the ΔcorA mutant produced MgtA, but not MgtB, in the presence of high extracellular magnesium concentration. Under these conditions, MgtA production in the ΔcorA mutant did not require PhoP and, consistently, a ΔmgtA, but not a ΔphoP, mutation slightly reduced the magnesium content of the ΔcorA mutant. Nevertheless, the ΔphoP mutation strongly impaired growth and motility, in the absence of CorA only and independently of the extracellular magnesium concentration, unraveling the importance of PhoP-dependent functions in the absence of CorA. Under these conditions, several proteins involved in flagella formation, chemotaxis and secretion were affected by the ΔcorA and ΔphoP mutations in combination, but not alone. Altogether, our data pinpoint a regulatory network, where the absence of CorA is sensed and its physiological consequences alleviated by induction of MgtA- and PhoP- dependent compensatory mechanisms.

Project description:Kaiser2014 - Salmonella persistence after ciprofloxacin treatment The model describes the bacterial tolerance to antibiotics. Using a mouse model for Salmonella diarrhea, the authors have found that bacterial persistence occurs in the presence of the antibiotic ciprofloxacin because Salmonella can exist in two different states. One, the fast-growing population that spreads in the host's tissues and the other, slow-growing "persister" population that hide out inside dendritic cells of the host's immune system and cannot be attacked by the antibiotics. However, this can be killed by adding agents that directly stimulate the host's immune defense. This model is described in the article: Cecum lymph node dendritic cells harbor slow-growing bacteria phenotypically tolerant to antibiotic treatment. Kaiser P, Regoes RR, Dolowschiak T, Wotzka SY, Lengefeld J, Slack E, Grant AJ, Ackermann M, Hardt WD. PLoS Biol. 2014 Feb 18;12(2):e1001793. Abstract: In vivo, antibiotics are often much less efficient than ex vivo and relapses can occur. The reasons for poor in vivo activity are still not completely understood. We have studied the fluoroquinolone antibiotic ciprofloxacin in an animal model for complicated Salmonellosis. High-dose ciprofloxacin treatment efficiently reduced pathogen loads in feces and most organs. However, the cecum draining lymph node (cLN), the gut tissue, and the spleen retained surviving bacteria. In cLN, approximately 10%-20% of the bacteria remained viable. These phenotypically tolerant bacteria lodged mostly within CD103⁺CX₃CR1⁻CD11c⁺ dendritic cells, remained genetically susceptible to ciprofloxacin, were sufficient to reinitiate infection after the end of the therapy, and displayed an extremely slow growth rate, as shown by mathematical analysis of infections with mixed inocula and segregative plasmid experiments. The slow growth was sufficient to explain recalcitrance to antibiotics treatment. Therefore, slow-growing antibiotic-tolerant bacteria lodged within dendritic cells can explain poor in vivo antibiotic activity and relapse. Administration of LPS or CpG, known elicitors of innate immune defense, reduced the loads of tolerant bacteria. Thus, manipulating innate immunity may augment the in vivo activity of antibiotics. This model is hosted on BioModels Database and identified by: MODEL1312170001. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. 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.

Dataset Information

Quantitative analysis of differential proteome expression in wild-type and mutants strains of Salmonella enterica serovar Typhimurium

Dataset's files

Publications

Proteome remodelling by the stress sigma factor RpoS/σ<sup>S</sup> in Salmonella: identification of small proteins and evidence for post-transcriptional regulation.

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