Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White

ABSTRACT The survival ability of Salmonella enterica serovar Enteritidis in antibacterial egg white is an important factor leading to Salmonella outbreaks through eggs and egg products. In this study, the role of the gene yoaE, encoding an inner membrane protein, in the survival of Salmonella Enteri...

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Autores principales:	Xiaozhen Huang, Mengjun Hu, Xiujuan Zhou, Yanhong Liu, Chunlei Shi, Xianming Shi
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	ultrafiltration matrix egg white yoaE cpxR alkaline pH antimicrobial peptide Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/cea6c30c4d3541ea975715c22e9dc4d3
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spelling	oai:doaj.org-article:cea6c30c4d3541ea975715c22e9dc4d32021-11-15T15:27:53ZRole of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White10.1128/mSphere.00638-192379-5042https://doaj.org/article/cea6c30c4d3541ea975715c22e9dc4d32020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00638-19https://doaj.org/toc/2379-5042ABSTRACT The survival ability of Salmonella enterica serovar Enteritidis in antibacterial egg white is an important factor leading to Salmonella outbreaks through eggs and egg products. In this study, the role of the gene yoaE, encoding an inner membrane protein, in the survival of Salmonella Enteritidis in egg white, and its transcriptional regulation by CpxR were investigated. Quantitative reverse transcription-PCR (RT-qPCR) results showed that the yoaE gene expression was upregulated 35-fold after exposure to egg white for 4 h compared to that in M9FeS medium, and the deletion of yoaE (ΔyoaE) dramatically decreased the survival rate of bacteria in egg white to less than 1% of the wild type (WT) and the complementary strain at both 37 and 20°C, indicating that yoaE was essential for bacteria to survive in egg white. Furthermore, the ΔyoaE strain was sensitive to a 3-kDa ultrafiltration matrix of egg white because of its high pH and antimicrobial peptide components. Putative conserved binding sites for the envelope stress response regulator CpxR were found in the yoaE promoter region. In vivo, the RT-qPCR assay results showed that the upregulation of yoaE in a ΔcpxR strain in egg white was 1/5 that of the WT. In vitro, results from DNase I footprinting and electrophoretic mobility shift assays further demonstrated that CpxR could directly bind to the yoaE promoter region, and a specific CpxR binding sequence was identified. In conclusion, it was shown for the first time that CpxR positively regulated the transcription of yoaE, which was indispensable for survival of Salmonella Enteritidis in egg white. IMPORTANCE Salmonella enterica serovar Enteritidis is the predominant Salmonella serotype that causes human salmonellosis mainly through contaminated chicken eggs or egg products and has been a global public health threat. The spread and frequent outbreaks of this serotype through eggs correlate significantly with its exceptional survival in eggs, despite the antibacterial properties of egg white. Research on the survival mechanisms of S. Enteritidis in egg white will help develop effective strategies to control the contamination of eggs by this Salmonella serotype and help further elucidate the complex antibacterial mechanisms of egg white. This study revealed the importance of yoaE, a gene with unknown function, on the survival of S. Enteritidis in egg white, as well as its transcriptional regulation by CpxR. Our work provides the basis to reveal the mechanisms of survival of S. Enteritidis in egg white and the specific function of the yoaE gene.Xiaozhen HuangMengjun HuXiujuan ZhouYanhong LiuChunlei ShiXianming ShiAmerican Society for Microbiologyarticleultrafiltration matrixegg whiteyoaEcpxRalkaline pHantimicrobial peptideMicrobiologyQR1-502ENmSphere, Vol 5, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	ultrafiltration matrix egg white yoaE cpxR alkaline pH antimicrobial peptide Microbiology QR1-502
spellingShingle	ultrafiltration matrix egg white yoaE cpxR alkaline pH antimicrobial peptide Microbiology QR1-502 Xiaozhen Huang Mengjun Hu Xiujuan Zhou Yanhong Liu Chunlei Shi Xianming Shi Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White
description	ABSTRACT The survival ability of Salmonella enterica serovar Enteritidis in antibacterial egg white is an important factor leading to Salmonella outbreaks through eggs and egg products. In this study, the role of the gene yoaE, encoding an inner membrane protein, in the survival of Salmonella Enteritidis in egg white, and its transcriptional regulation by CpxR were investigated. Quantitative reverse transcription-PCR (RT-qPCR) results showed that the yoaE gene expression was upregulated 35-fold after exposure to egg white for 4 h compared to that in M9FeS medium, and the deletion of yoaE (ΔyoaE) dramatically decreased the survival rate of bacteria in egg white to less than 1% of the wild type (WT) and the complementary strain at both 37 and 20°C, indicating that yoaE was essential for bacteria to survive in egg white. Furthermore, the ΔyoaE strain was sensitive to a 3-kDa ultrafiltration matrix of egg white because of its high pH and antimicrobial peptide components. Putative conserved binding sites for the envelope stress response regulator CpxR were found in the yoaE promoter region. In vivo, the RT-qPCR assay results showed that the upregulation of yoaE in a ΔcpxR strain in egg white was 1/5 that of the WT. In vitro, results from DNase I footprinting and electrophoretic mobility shift assays further demonstrated that CpxR could directly bind to the yoaE promoter region, and a specific CpxR binding sequence was identified. In conclusion, it was shown for the first time that CpxR positively regulated the transcription of yoaE, which was indispensable for survival of Salmonella Enteritidis in egg white. IMPORTANCE Salmonella enterica serovar Enteritidis is the predominant Salmonella serotype that causes human salmonellosis mainly through contaminated chicken eggs or egg products and has been a global public health threat. The spread and frequent outbreaks of this serotype through eggs correlate significantly with its exceptional survival in eggs, despite the antibacterial properties of egg white. Research on the survival mechanisms of S. Enteritidis in egg white will help develop effective strategies to control the contamination of eggs by this Salmonella serotype and help further elucidate the complex antibacterial mechanisms of egg white. This study revealed the importance of yoaE, a gene with unknown function, on the survival of S. Enteritidis in egg white, as well as its transcriptional regulation by CpxR. Our work provides the basis to reveal the mechanisms of survival of S. Enteritidis in egg white and the specific function of the yoaE gene.
format	article
author	Xiaozhen Huang Mengjun Hu Xiujuan Zhou Yanhong Liu Chunlei Shi Xianming Shi
author_facet	Xiaozhen Huang Mengjun Hu Xiujuan Zhou Yanhong Liu Chunlei Shi Xianming Shi
author_sort	Xiaozhen Huang
title	Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White
title_short	Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White
title_full	Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White
title_fullStr	Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White
title_full_unstemmed	Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White
title_sort	role of <italic toggle="yes">yoae</italic> gene regulated by cpxr in the survival of <italic toggle="yes">salmonella enterica</italic> serovar enteritidis in antibacterial egg white
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/cea6c30c4d3541ea975715c22e9dc4d3
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Role of <italic toggle="yes">yoaE</italic> Gene Regulated by CpxR in the Survival of <italic toggle="yes">Salmonella enterica</italic> Serovar Enteritidis in Antibacterial Egg White

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