The Aeromonas salmonicida subsp. salmonicida exoproteome: global analysis, moonlighting proteins and putative antigens for vaccination against furunculosis

Abstract

BackgroundAeromonas salmonicida subsp. salmonicida, the etiologic agent of furunculosis, is a major pathogen of fisheries worldwide. Despite the identification of several virulence factors the pathogenesis is still poorly understood. We have used high-throughput proteomics to display the differences between in vitro secretome of A. salmonicida wild-type (wt, hypervirulent, JF5054) and T3SS-deficient (isogenic ΔascV, extremely low-virulent, JF2747) strains in exponential (GP) and stationary (SP) phases of growth.ResultsAmong the different experimental conditions we obtained semi-quantitative values for a total of 2136 A. salmonicida proteins. Proteins of specific A. salmonicida species were proportionally less detected than proteins common to the Aeromonas genus or those shared with other Aeromonas species, suggesting that in vitro growth did not induce the expression of these genes. Four detected proteins which are unidentified in the genome of reference strains of A. salmonicida were homologous to components of the conjugative T4SS of A. hydrophila pRA1 plasmid. Polypeptides of three proteins which are specific to the 01-B526 strain were also discovered. In supernatants (SNs), the number of detected proteins was higher in SP (326 for wt vs 329 for mutant) than in GP (275 for wt vs 263 for mutant). In pellets, the number of identified proteins (a total of 1536) was approximately the same between GP and SP. Numerous highly conserved cytoplasmic proteins were present in A. salmonicida SNs (mainly EF-Tu, EF-G, EF-P, EF-Ts, TypA, AlaS, ribosomal proteins, HtpG, DnaK, peptidyl-prolyl cis-trans isomerases, GAPDH, Enolase, FbaA, TpiA, Pgk, TktA, AckA, AcnB, Mdh, AhpC, Tpx, SodB and PNPase), and several evidences support the theory that their extracellular localization was not the result of cell lysis. According to the Cluster of Orthologous Groups classification, 29% of excreted proteins in A. salmonicida SNs were currently poorly characterized.ConclusionsIn this part of our work we elucidated the whole in vitro exoproteome of hypervirulent A. salmonicida subsp. salmonicida and showed the secretion of several highly conserved cytoplasmic proteins with putative moonlighting functions and roles in virulence. All together, our results offer new information about the pathogenesis of furunculosis and point out potential candidates for vaccine development.

Type III Secretion System of Aeromonas salmonicida Undermining the Host's Immune Response

“…AopN is involved in controlling the secretion of translocator proteins as a gate keeper inside the bacterium. However, it is also secreted by the T3SS in virulent A. salmonicida like effector proteins (Vanden Bergh et al, 2013b). Secretion of AopN, however occurred at a somewhat lower extent than the other effectors described (Bergh et al, 2013).…”

“…The role of AopH in A. salmonicida has not yet been studied by itself. However, it is thought to have a prominent activity in disturbing cell signaling that would be responsible for the initiation of the adaptive immune response (Vanden Bergh et al, 2013b;Vanden Bergh and Frey, 2014).…”

Missing the target: DNAk is a dominant epitope in the humoral immune response of channel catfish (Ictalurus punctatus) to Flavobacterium columnare

“…Their identification was among a long list of cellular proteins that are "moonlighting" in the ECP, and they suggested perhaps that DNAk would be a viable vaccine candidate (Vanden Bergh et al, 2013). We also show that 8E2/2 and more importantly catfish antiserum generated against F. columnare are reactive to the DNAk proteins of several gram negative bacterial fish pathogens.…”

Antigens of Aeromonas salmonicida subsp. salmonicida specifically induced in vivo in Oncorhynchus mykiss

“… Vanden Bergh et al . ), a virulence mechanism that is considered particularly important in A. salmonicida (Burr et al . ).…”