Abstract

Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia, a chronic respiratory disease in swine resulting in enormous economic losses. To identify the components that contribute to virulence and unveil those biological processes potentially related to attenuation, we used isobaric tags for relative and absolute quantification technology (iTRAQ) to compare the protein profiles of the virulent M. hyopneumoniae strain 168 and its attenuated highly passaged strain 168L. We identified 489 proteins in total, 70 of which showing significant differences in level of expression between the two strains. Remarkably, proteins participating in inositol phosphate metabolism were significantly downregulated in the virulent strain, while some proteins involved in nucleoside metabolism were upregulated. We also mined a series of novel promising virulence-associated factors in our study compared with those in previous reports, such as some moonlighting adhesins, transporters, lipoate-protein ligase, and ribonuclease and several hypothetical proteins with conserved functional domains, deserving further research. Our survey constitutes an iTRAQ-based comparative proteomic analysis of a virulent M. hyopneumoniae strain and its attenuated strain originating from a single parent with a well-characterized genetic background and lays the groundwork for future work to mine for potential virulence factors and identify candidate vaccine proteins.

Highlights

  • M. hyopneumoniae is the causative agent of porcine enzootic pneumonia, which is a worldwide epidemic that can cause enormous economic losses as a result of retarded growth in pigs and the cost of disease control [1]

  • Of the 489 proteins identified, 273 proteins were annotated into 9 groups based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) categories

  • We found that in our data four enzymes participated in nucleoside metabolism: purinenucleoside phosphorylase (DeoD), hypoxanthine-guanine phosphoribosyl transferase (Hpt), thymidine kinase (Tdk), and CTP synthase (PyrG), all upregulated in the virulent strain

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Summary

Introduction

M. hyopneumoniae is the causative agent of porcine enzootic pneumonia, which is a worldwide epidemic that can cause enormous economic losses as a result of retarded growth in pigs and the cost of disease control [1]. Despite its low direct mortality, M. hyopneumoniae increases the host’s susceptibility to secondary respiratory infections by damaging cilia and epithelial cell, resulting in aggravated lung lesions and fatal respiratory diseases [2]. The determination of the potential molecular mechanisms of the pathogenicity of M. hyopneumoniae has been hampered by the fastidious growth requirements of this organism and the lack of tools for its genetic manipulation. Comparative genomic analyses have been performed to reveal the genetic basis of virulence attenuation of M. hyopneumoniae and to predict potential virulence factors [3,4,5]. We decided to turn our attention to the variations in expression levels of the predicted virulence factors

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