Interactome of E. piscicida and grouper liver proteins reveals strategies of bacterial infection and host immune response

Abstract

The occurrence of infectious diseases is related to heterogeneous protein interactions between a host and a microbe. Therefore, elucidating the host-pathogen interplay is essential. We previously revealed the protein interactome between Edwardsiella piscicida and fish gill cells, and the present study identified the protein interactome between E. piscicida and E. drummondhayi liver cells. E. drummondhayi liver cells and bacterial pull-down approaches were used to identify E. piscicida outer membrane proteins that bind to liver cells and fish liver cell proteins that interact with bacterial cells, respectively. Eight bacterial proteins and 11 fish proteins were characterized. Heterogeneous protein-protein interactions between these bacterial cells and fish liver cells were investigated through far-Western blotting and co-immunoprecipitation. A network was constructed based on 42 heterogeneous protein-protein interactions between seven bacterial proteins and 10 fish proteins. A comparison of the new interactome with the previously reported interactome showed that four bacterial proteins overlapped, whereas all of the identified fish proteins were new, suggesting a difference between bacterial tricks for evading host immunity and the host strategy for combating bacterial infection. Furthermore, these bacterial proteins were found to regulate the expression of host innate immune-related proteins. These findings indicate that the interactome contributes to bacterial infection and host immunity.

Edwardsiella piscicida: A versatile emerging pathogen of fish

“…These include proteomics and genomics studies as well as secretomes, transcriptomes, interactomes, and metabolomics studies. Proteomics and metabolomics studies have enabled researchers to examine protein-protein interactions between E. piscicida cells and host tissues, such as fish gills [37] and livers [106]; in antibiotics resistome variations [107] and stress adaptation [108]. RNA-seq, ChIP-seq and Tn-seq are new and powerful technologies that couple defined transposon mutant library with high-throughput sequencing of transposon insertion sites to comprehensively map genetic determinants of bacterial fitness.…”

Protein–protein interaction network: an emerging tool for understanding fish disease in aquaculture

“…Diseases are often indicated or caused by the disruptions (mutations and defects) of protein complexes, resulting in adverse changes to the normal pattern of PPIs (Hao et al 2014;Li et al 2017). Although some disruptions might not have significant functional consequences, other changes directly lead to the initiation or progression of a disease.…”

“…Proteins interact with each other and form complexes to ensure the proper functioning of normal cellular processes. Any perturbations that occur to a specific protein will alter the normal pattern of PPI, thus contributing to a disease state (Hao et al 2014;Li et al 2017). Therefore, PPI networks are gaining attention in the fields of human pathobiology, and plant physiology and pathology.…”

The LysR-Type Transcriptional Regulator YeeY Plays Important Roles in the Regulatory of Furazolidone Resistance in Aeromonas hydrophila

“…Briefly, the pET-32a plasmid containing yeeY was transformed into E. coli BL21 (DE3). The recombinant proteins were induced with IPTG (isopropyl β-D-1-thiogalactopyranoside, 0.05 mM) at 30 • C for 6 h and then purified using Ni-TNA column as described in previous study (Li et al, 2017). Second, two DNA probes were synthesized by PCR using two pairs of single-strand primers (Table 1) in which the upstream primer was labeled by Cy5-labeled oligonucleotides at the 5 end, whereas two cold probes as a competitor were synthesized with the primers without the Cy5-labeled at the upstream primer 5 end ( Table 1).…”