Dataset Information

ITRAQ-based quantitative proteomic analysis reveals potential virulence factors of Erysipelothrix rhusiopathiae

ABSTRACT: Erysipelothrix rhusiopathiae is a small, facultatively aerobic, gram-positive bacterium that causes erysipelas in swine and a wide spectrum of diseases in other animals, as well as the skin disease erysipeloid in humans. Swine erysipelas caused by E. rhusiopathiae is the most prevalent and economically important of these diseases. In China, swine erysipelas has emerged at an alarming rate, developing from scattered occurrences on a small number of farms to systemic outbreaks since 2012 in many provinces, thereby inflicting large financial losses. Some virulence factors of E. rhusiopathiae have been identified. The capsule is considered essential for resistance to phagocytosis and bacterial virulence. Neuraminidase plays a crucial role in initiating adhesion to endothelial cells. Two surface-adhesion proteins (RspA and RspB) participate in the initiation of biofilm formation by binding to abiotic and biotic surfaces. SpaA is involved in the adhesion of E. rhusiopathiae to porcine endothelial cells. Nevertheless, very little is known about the pathogenesis of E. rhusiopathiae. This knowledge gap has become one of the most important obstacles to controlling infection. Thus, the identification of novel virulence factors is necessary to improve understanding of the pathogenesis of E. rhusiopathiae.Surface proteins of gram-positive bacteria play a critical role in virulence by modifying the environment surrounding the cell. Furthermore, for many bacteria, surface proteins are candidate antigens for subunit vaccines. The examination of bacterial surface-protein fractions, including cell wall-associated proteins (CWPs), is an effective method for identifying novel virulence factors. One proteomic approach involves isobaric Tags for Relative and Absolute Quantitation (iTRAQ) coupled to liquid chromatography (LC)-quadrupole mass spectrometry (MS/MS). This method has been widely used for the comparative quantitative analysis of bacterial proteomes. In this study, iTRAQ combined with LC-MS/MS was applied to compare differentially abundant CWPs of high- and low-virulence strains of E. rhusiopathiae. Proteins with higher abundance in the high-virulence strain were found to aid in predicting potential virulence factors to improve vaccines or disease-control strategies.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Erysipelothrix Rhusiopathiae

SUBMITTER: Ya Wang

LAB HEAD: Meilin Jin

PROVIDER: PXD005505 | Pride | 2017-03-28

REPOSITORIES: Pride

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iTRAQ-based quantitative proteomic analysis reveals potential virulence factors of Erysipelothrix rhusiopathiae.

Wang Ya Y Li Jingtao J Zhang Anding A Zhu Weifeng W Zhang Qiang Q Xu Zhongmin Z Yan Shuxian S Sun Xiaomei X Chen Huanchun H Jin Meilin M

Journal of proteomics 20170308

Erysipelothrix rhusiopathiae is a ubiquitous pathogen that has caused considerable economic losses to pig farmers. However, the mechanisms of E. rhusiopathiae pathogenesis remain unclear. To identify new virulence-associated factors, the differentially abundant cell wall-associated proteins (CWPs) between high- and low-virulence strains were investigated through isobaric Tags for Relative and Absolute Quantitation (iTRAQ) combined with liquid chromatography-quadrupole mass spectrometry (LC-MS/MS ...[more]

PMID: 28284755

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Project description:Enteric pathogens have developed several resistance mechanisms to survive the antimicrobial action of bile. We investigated the transcriptional profile of Vibrio cholerae O1 El Tor strain C6706 under virulence gene inducing conditions, in the presence and absence of bile. Microarray analysis revealed that the expression of 119 genes was affected by bile. The mRNA levels of genes encoding proteins involved in transport was increased in the presence of bile, whereas mRNA levels of genes encoding proteins involved in pathogenesis and chemotaxis was decreased. This study identified genes encoding transcriptional regulators from the TetR family (vexR and breR) and multidrug efflux pumps from the RND superfamily (vexB and vexD [here renamed breB]) that were induced in response to bile. Further analysis regarding vexAB and breAB expression in the presence of various antimicrobial compounds established that vexAB was induced in the presence of bile, SDS or novobiocin, whereas induction of breAB was specific to bile. BreR is a direct repressor of the breAB promoter and is able to autoregulate its own expression, as demonstrated by transcriptional and electrophoretic mobility shift assays (EMSA). Expression of breR and breAB is induced in the presence of the bile salts cholate, deoxycholate or chenodeoxycholate and EMSA showed that deoxycholate is able to abolish formation of BreR-PbreR complexes. We propose that deoxycholate is able to interact with BreR and induce a conformational change that will interfere with its DNA binding ability resulting in breAB and breR expression. These results provide new insight into a transcriptional regulator and a transport system that likely play an essential role in the ability of Vibrio cholerae to resist the action of bile in the host. Keywords: Vibrio cholerae response to bile Three independent experiments were performed for the microarray analysis. From each experiment, RNA was obtained from four different time points and was prepared for hybridization, therefore, a total of 3 slides were analyzed per time point. The growth conditions were as follows; C6706 str2 was grown 15 h in LB medium at 30 degrees C with aeration. The cultures were then diluted 100-fold into AKI medium with, or without, 0.4% crude bile (high bile concentration) and were grown at 37degrees C for 3.5 h stationary followed by 2 h with aeration to induce virulence gene expression. Then the cultures were diluted 100-fold into AKI medium with, or without, 0.02% crude bile (low bile concentration) and were grown at 37 degrees C for 2 h stationary. Samples were obtained from 4 different time points: 2, 4, and 5.5 h from the high bile cultures, and at 2 h from the low bile cultures.

Dataset Information

ITRAQ-based quantitative proteomic analysis reveals potential virulence factors of Erysipelothrix rhusiopathiae

Publications

iTRAQ-based quantitative proteomic analysis reveals potential virulence factors of Erysipelothrix rhusiopathiae.

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