Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen

ABSTRACT Commensal and generally harmless in healthy individuals, Enterococcus faecalis causes opportunistic infections in immunocompromised patients. Plasmid-cured E. faecalis strain VE14089, derived from sequenced reference strain V583, is widely used for functional studies due to its improved gen...

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Autores principales:	Sylviane Furlan, Renata C. Matos, Sean P. Kennedy, Benoît Doublet, Pascale Serror, Lionel Rigottier-Gois
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Enterococcus faecalis epa variable region genome and SNP analyses growth fitness host adaptation platform strain Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/64e25b8e564244038f80cf8742cdefcc
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spelling	oai:doaj.org-article:64e25b8e564244038f80cf8742cdefcc2021-11-15T15:22:26ZFitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen10.1128/mSphere.00310-192379-5042https://doaj.org/article/64e25b8e564244038f80cf8742cdefcc2019-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00310-19https://doaj.org/toc/2379-5042ABSTRACT Commensal and generally harmless in healthy individuals, Enterococcus faecalis causes opportunistic infections in immunocompromised patients. Plasmid-cured E. faecalis strain VE14089, derived from sequenced reference strain V583, is widely used for functional studies due to its improved genetic amenability. Although strain VE14089 has no major DNA rearrangements, with the exception of an ∼20-kb integrated region of pTEF1 plasmid, the strain presented significant growth differences from the V583 reference strain of our collection (renamed VE14002). In the present study, genome sequencing of strain VE14089 identified additional point mutations. Excision of the integrated pTEF1 plasmid region and sequential restoration of wild-type alleles showing nonsilent mutations were performed to obtain the VE18379 reference-derivative strain. Recovery of the growth ability of the restored VE18379 strain at a level similar to that seen with the reference strain points to GreA and Spx as bacterial fitness determinants. Virulence potential in Galleria mellonella and intestinal colonization in mouse demonstrated host adaptation of the VE18379 strain equivalent to VE14002 host adaptation. We further demonstrated that deletion of the 16.8-kb variable region of the epa locus recapitulates the key role of Epa decoration in host adaptation, providing a genetic system to study the role of specific epa-variable regions in host adaptation independently of other genetic variations. IMPORTANCE E. faecalis strain VE14089 was derived from V583 cured of its plasmids. Although VE14089 had no major DNA rearrangements, it presented significant growth and host adaptation differences from the reference strain V583 of our collection. To construct a strain with better fitness, we sequenced the genome of VE14089, identified single nucleotide polymorphisms (SNPs), and repaired the genes that could account for these changes. Using this reference-derivative strain, we provide a novel genetic system to understand the role of the variable region of epa in the enterococcal lifestyle.Sylviane FurlanRenata C. MatosSean P. KennedyBenoît DoubletPascale SerrorLionel Rigottier-GoisAmerican Society for MicrobiologyarticleEnterococcus faecalisepa variable regiongenome and SNP analysesgrowth fitnesshost adaptationplatform strainMicrobiologyQR1-502ENmSphere, Vol 4, Iss 4 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Enterococcus faecalis epa variable region genome and SNP analyses growth fitness host adaptation platform strain Microbiology QR1-502
spellingShingle	Enterococcus faecalis epa variable region genome and SNP analyses growth fitness host adaptation platform strain Microbiology QR1-502 Sylviane Furlan Renata C. Matos Sean P. Kennedy Benoît Doublet Pascale Serror Lionel Rigottier-Gois Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen
description	ABSTRACT Commensal and generally harmless in healthy individuals, Enterococcus faecalis causes opportunistic infections in immunocompromised patients. Plasmid-cured E. faecalis strain VE14089, derived from sequenced reference strain V583, is widely used for functional studies due to its improved genetic amenability. Although strain VE14089 has no major DNA rearrangements, with the exception of an ∼20-kb integrated region of pTEF1 plasmid, the strain presented significant growth differences from the V583 reference strain of our collection (renamed VE14002). In the present study, genome sequencing of strain VE14089 identified additional point mutations. Excision of the integrated pTEF1 plasmid region and sequential restoration of wild-type alleles showing nonsilent mutations were performed to obtain the VE18379 reference-derivative strain. Recovery of the growth ability of the restored VE18379 strain at a level similar to that seen with the reference strain points to GreA and Spx as bacterial fitness determinants. Virulence potential in Galleria mellonella and intestinal colonization in mouse demonstrated host adaptation of the VE18379 strain equivalent to VE14002 host adaptation. We further demonstrated that deletion of the 16.8-kb variable region of the epa locus recapitulates the key role of Epa decoration in host adaptation, providing a genetic system to study the role of specific epa-variable regions in host adaptation independently of other genetic variations. IMPORTANCE E. faecalis strain VE14089 was derived from V583 cured of its plasmids. Although VE14089 had no major DNA rearrangements, it presented significant growth and host adaptation differences from the reference strain V583 of our collection. To construct a strain with better fitness, we sequenced the genome of VE14089, identified single nucleotide polymorphisms (SNPs), and repaired the genes that could account for these changes. Using this reference-derivative strain, we provide a novel genetic system to understand the role of the variable region of epa in the enterococcal lifestyle.
format	article
author	Sylviane Furlan Renata C. Matos Sean P. Kennedy Benoît Doublet Pascale Serror Lionel Rigottier-Gois
author_facet	Sylviane Furlan Renata C. Matos Sean P. Kennedy Benoît Doublet Pascale Serror Lionel Rigottier-Gois
author_sort	Sylviane Furlan
title	Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen
title_short	Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen
title_full	Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen
title_fullStr	Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen
title_full_unstemmed	Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen
title_sort	fitness restoration of a genetically tractable <named-content content-type="genus-species">enterococcus faecalis</named-content> v583 derivative to study decoration-related phenotypes of the enterococcal polysaccharide antigen
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/64e25b8e564244038f80cf8742cdefcc
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Fitness Restoration of a Genetically Tractable <named-content content-type="genus-species">Enterococcus faecalis</named-content> V583 Derivative To Study Decoration-Related Phenotypes of the Enterococcal Polysaccharide Antigen

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