Direct interactions between bidensovirus BmDNV-Z proteins and midgut proteins from the virus target Bombyx mori

In this study we aimed to identify the protein-protein interactions between Bombyx mori midgut and the bidensovirus BmDNV-Z via a yeast two-hybrid (Y2H) system. To achieve this we constructed a Gal4 activation domain fusion library that expresses the host genes and Gal4 DNA binding domain fusion bait vectors that express BmDNV-Z genes. Y2H assay revealed 15 potential interactions between host and viral proteins. To verify the interactions, we modified and reconstructed a pair of bimolecular fluorescence complementation (BiFC) vectors and achieved the co-expressions of the candidate host genes and viral genes in insect culture cells. The BiFC assay confirmed the specificity of the interactions including B. mori 35 kDa protease and two BmDNV-Z proteins encoded by VD1-ORF2 and VD2-ORF1; B. mori transgelin and BmDNV-Z protein encoded by VD2-ORF3; and B. mori serine protease precursor and BmDNV-Z encoded by VD2-ORF3 in vitro. Our findings revealed that the specific host midgut proteins are involved in the interactions between B. mori and BmDNV-Z, which will facilitate our understanding of the molecular mechanisms of BmDNV-Z infection.