Construction and application of a protein interaction map for white spot syndrome virus (WSSV).
White spot syndrome virus (WSSV) is currently the most serious global threat for cultured shrimp production. Although its large, double-stranded DNA genome has been completely characterized, most putative protein functions remain obscure. To provide more informative knowledge about this virus, a proteomic-scale network of WSSV-WSSV protein interactions was carried out using a comprehensive yeast two-hybrid analysis. An array of yeast transformants containing each WSSV open reading frame fused with GAL4 DNA binding domain and GAL4 activation domain was constructed yielding 187 bait and 182 prey constructs, respectively. On screening of ∼28,000 pairwise combinations, 710 interactions were obtained from 143 baits. An independent coimmunoprecipitation assay (co-IP) was performed to validate the selected protein interaction pairs identified from the yeast two-hybrid approach. The program Cytoscape was employed to create a WSSV protein-protein interaction (PPI) network. The topology of the WSSV PPI network was based on the Barabási-Albert model and consisted of a scale-free network that resembled other established viral protein interaction networks. Using the RNA interference approach, knocking down either of two candidate hub proteins gave shrimp more protection against WSSV than knocking down a nonhub gene. The WSSV protein interaction map established in this study provides novel guidance for further studies on shrimp viral pathogenesis, host-viral protein interaction and potential targets for therapeutic and preventative antiviral strategies in shrimp aquaculture.
Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand;
Pakkakul
P
Sangsuriya
Jiun-Yan
JY
Huang
Yu-Fei
YF
Chu
Kornsunee
K
Phiwsaiya
Pimlapas
P
Leekitcharoenphon
Watcharachai
W
Meemetta
Saengchan
S
Senapin
Wei-Pang
WP
Huang
Boonsirm
B
Withyachumnarnkul
Timothy W
TW
Flegel
Chu-Fang
CF
Lo
D000818:Animals; D054884:Host-Pathogen Interactions / Q000235:genetics; D033561:Penaeidae / Q000821:virology*; D060066:Protein Interaction Maps / Q000235:genetics*; D040901:Proteomics; D059467:Transcriptome; D014764:Viral Proteins / Q000235:genetics; D046848:White spot syndrome virus 1 / Q000235:genetics* / Q000378:metabolism
D014764:Viral Proteins