Mycoplasma synoviae lipid-associated membrane proteins identification and expression changes when exposed to chicken cells

Abstract

Mycoplasma synoviae is a significant cause of respiratory disease and synovitis among chickens, and has an adverse economic impact on broiler breeding efforts. The present study was designed to develop a systematic understanding of the role that M. synoviae lipid-associated membrane proteins (LAMPs) may play in the virulence of this pathogen. Bioinformatics tools were used to identify 146 predicted membrane proteins and lipoproteins in the M. synoviae proteome. Then, Triton X-114 was used to extract LAMPs that were subsequently identified via LC–MS/MS. This approach enabled the detection of potential LAMPs, and the top 200 most abundant proteins detected using this strategy were subject to further analysis. M. synoviae cells (100 MOI) were exposed to chicken fibroblasts (DF-1) and macrophages (HD-11) in a 1:1 mixed culture. Analysis of LAMP transcripts identified 72 up-regulated LAMP genes which were analyzed in depth by bioinformatics. GO analysis revealed these genes to be enriched in the nucleotide binding, sulfur amino acid transmembrane transporter activity, tRNA binding, rRNA modification, and transition metal ion transport pathways. Moreover, KEGG enrichment analysis suggested that these genes were enriched in the biosynthesis of secondary metabolites, carbon metabolism, glycolysis/gluconeogenesis, and nitrogen metabolism pathways.

Hypothetical Proteins of Mycoplasma synoviae Reannotation and Expression Changes Identified via RNA-Sequencing

“…The subcellular localization predictions indicated that M. synoviae HPs would be found in the cytoplasm, cytoplasmic membrane, or extracellular fractions. Some HPs predicted to be both cytoplasmic and extracellular proteins were found in the surface fraction in our previous proteomic study [38]. Therefore, some HPs were also located in multiple compartments and/or served moonlight functions [42].…”

“…The M. synoviae ATCC 25204 genome consists of 720 CDSs and 673 genes, among which 155 encode HPs. However, only 8.39% of all HPs (13 out of 155) have been detected in membrane-associated lipoprotein proteomic studies [38].…”