Cold-shock RNA-binding protein CspR is also exposed to the surface of 
               
                  Enterococcus faecalis

Michaux, Charlotte; Saavedra, Luis Felipe Romero; Reffuveille, Fany; Bernay, Benoı̂t; Goux, Didier; Hartke, Axel; Verneuil, Nicolas; Giard, Jean‐Christophe

doi:10.1099/mic.0.071076-0

Cited by 9 publications

(5 citation statements)

References 54 publications

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“…Four of the seven proteins that were predicted to be located intracellularly by LocateP (transmembrane segment absent) but that were identified after proteolytic shaving were recently also identified at the cell surface in other bacterial species. In Enterococcus faecalis proteolytic shaving revealed the presence of cold shock proteins (Csp) at the cell surface, which was confirmed by electron microscopy using anti‐Csp antibodies . In our analysis, we identified two proteins after growth in urine and shaving, EFME1162_2006 and EFME1162_0442, which were also annotated as cold shock proteins.…”

Section: Resultssupporting

confidence: 57%

Growth condition‐dependent cell surface proteome analysis of Enterococcus faecium

et al. 2015

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The last 30 years Enterococcus faecium has become an important nosocomial pathogen in hospitals worldwide. The aim of this study was to obtain insight in the cell surface proteome of E. faecium when grown in laboratory and clinically relevant conditions. Enterococcus faecium E1162, a clinical blood stream isolate, was grown until mid-log phase in brain heart infusion medium (BHI) with, or without 0.02% bile salts, Tryptic Soy Broth with 1% glucose (TSBg) and urine, and its cell surface was "shaved" using immobilized trypsin. Peptides were identified using MS/MS. Mapping against the translated E1162 whole genome sequence identified 67 proteins that were differentially detected in different conditions. In urine, 14 proteins were significantly more and nine proteins less abundant relative to the other conditions. Growth in BHI-bile and TSBg, revealed four and six proteins, respectively, which were uniquely present in these conditions while two proteins were uniquely present in both conditions. Thus, proteolytic shaving of E. faecium cells identified differentially surface exposed proteins in different growth conditions. These proteins are of special interest as they provide more insight in the adaptive mechanisms and may serve as targets for the development of novel therapeutics against this multi-resistant emerging pathogen. All MS data have been deposited in the ProteomeXchange with identifier PXD002497 (http://proteomecentral.proteomexchange.org/dataset/PXD002497).

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Section: Resultssupporting

confidence: 57%

Growth condition‐dependent cell surface proteome analysis of Enterococcus faecium

et al. 2015

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“…This identified 189 genes as supported by stringent statistical measures. However, previously proven or likely false negatives remained in the candidate set, including DltC (56) and CspA (57,58). Therefore, an additional 28 genes were added to the list of resistance determinants based on an antibiotic challenge dVal of Ͻ0.100 and manual inspection of data and literature support, creating a set of 217 genes most likely to contribute to intrinsic antibiotic resistance in E. faecalis MMH594 (Data Set S8a).…”

Section: Resultsmentioning

confidence: 99%

Genes Contributing to the Unique Biology and Intrinsic Antibiotic Resistance of Enterococcus faecalis

Gilmore

Salamzade

Selleck

et al. 2020

mBio

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The enterococci, which are among the leading causes of multidrug-resistant (MDR) hospital infection, are notable for their environmental ruggedness, which extends to intrinsic antibiotic resistance. To identify genes that confer this unique property, we used Tn-seq to comprehensively explore the genome of MDR Enterococcus faecalis strain MMH594 for genes important for growth in nutrient-containing medium and with low-level antibiotic challenge. As expected, a large core of genes for DNA replication, expression, and central metabolism, shared with other bacteria, are intolerant to transposon disruption. However, genes were identified that are important to E. faecalis that are either absent from or unimportant for Staphylococcus aureus and Streptococcus pneumoniae fitness when similarly tested. Further, 217 genes were identified that when challenged by sub-MIC antibiotic levels exhibited reduced tolerance to transposon disruption, including those previously shown to contribute to intrinsic resistance, and others not previously ascribed this role. E. faecalis is one of the few Gram-positive bacteria experimentally shown to possess a functional Entner-Doudoroff pathway for carbon metabolism, a pathway that contributes to stress tolerance in other microbes. Through functional genomics and network analysis we defined the unusual structure of this pathway in E. faecalis and assessed its importance. These approaches also identified toxin-antitoxin and related systems that are unique and active in E. faecalis. Finally, we identified genes that are absent in the closest nonenterococcal relatives, the vagococci, and that contribute importantly to fitness with and without antibiotic selection, advancing an understanding of the unique biology of enterococci. IMPORTANCE Enterococci are leading causes of antibiotic-resistant infection transmitted in hospitals. The intrinsic hardiness of these organisms allows them to survive disinfection practices and then proliferate in the gastrointestinal tracts of antibiotic-treated patients. The objective of this study was to identify the underlying genetic basis for its unusual hardiness. Using a functional genomic approach, we identified traits and pathways of general importance for enterococcal survival and growth that distinguish them from closely related pathogens as well as ancestrally related species. We further identified unique traits that enable them to survive antibiotic challenge, revealing a large set of genes that contribute to intrinsic antibiotic resistance and a smaller set of uniquely important genes that are rare outside enterococci.

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“…Surface proteins of P. freudenreichii ITG P20 [ Centre International de Ressources Microbiennes-Bactéries d’Intérêt Alimentaire (CIRM-BIA) 129], which is used as a cheese ripening starter ( Richoux et al, 1998 ; Thierry et al, 2004 ), were investigated by a combination of proteomic methods previously developed for bacteria and eukaryotic cells ( Lortal et al, 1993 ; Mayrhofer et al, 2006 ; Rodríguez-Ortega et al, 2006 ; Berlec et al, 2011 ; Bøhle et al, 2011 ; Bensi et al, 2012 ; Ythier et al, 2012 ; Michaux et al, 2013 ). This investigation demonstrated the involvement of certain S-layer proteins in immunomodulation ( Bryson et al, 2006 ; Le Maréchal et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Propionibacterium freudenreichii Surface Protein SlpB Is Involved in Adhesion to Intestinal HT-29 Cells

et al. 2017

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Propionibacterium freudenreichii is a beneficial bacterium traditionally used as a cheese ripening starter and more recently for its probiotic abilities based on the release of beneficial metabolites. In addition to these metabolites (short-chain fatty acids, vitamins, and bifidogenic factor), P. freudenreichii revealed an immunomodulatory effect confirmed in vivo by the ability to protect mice from induced acute colitis. This effect is, however, highly strain-dependent. Local action of metabolites and of immunomodulatory molecules is favored by the ability of probiotics to adhere to the host cells. This property depends on key surface compounds, still poorly characterized in propionibacteria. In the present study, we showed different adhesion rates to cultured human intestinal cells, among strains of P. freudenreichii. The most adhesive one was P. freudenreichii CIRM-BIA 129, which is known to expose surface-layer proteins. We evidenced here the involvement of these proteins in adhesion to cultured human colon cells. We then aimed at deciphering the mechanisms involved in adhesion. Adhesion was inhibited by antibodies raised against SlpB, one of the surface-layer proteins in P. freudenreichii CIRM-BIA 129. Inactivation of the corresponding gene suppressed adhesion, further evidencing the key role of slpB product in cell adhesion. This work confirms the various functions fulfilled by surface-layer proteins, including probiotic/host interactions. It opens new perspectives for the understanding of probiotic determinants in propionibacteria, and for the selection of the most efficient strains within the P. freudenreichii species.

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Cold-shock RNA-binding protein CspR is also exposed to the surface of Enterococcus faecalis

Cited by 9 publications

References 54 publications

Growth condition‐dependent cell surface proteome analysis of Enterococcus faecium

Growth condition‐dependent cell surface proteome analysis of Enterococcus faecium

Genes Contributing to the Unique Biology and Intrinsic Antibiotic Resistance of Enterococcus faecalis

Propionibacterium freudenreichii Surface Protein SlpB Is Involved in Adhesion to Intestinal HT-29 Cells

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