Direct interactions between bidensovirus B m DNV ‐ Z proteins and midgut proteins from the virus target B ombyx mori

作者：Yan‐Yuan Bao, Libo Chen, Wenjuan Wu, Dong Zhao, Ying Wang, Xia Qin, Chuan‐Xi Zhang · 发表于：FEBS Journal · 年份：2012 · DOI：10.1111/febs.12088 · 被引用次数：18 · 研究领域：Viral Infectious Diseases and Gene Expression in Insects、Mosquito-borne diseases and control、Invertebrate Immune Response Mechanisms

In this study we aimed to identify the protein–protein interactions between B ombyx mori midgut and the bidensovirus B m DNV ‐ Z via a yeast two‐hybrid ( Y 2 H ) system. To achieve this we constructed a G al4 activation domain fusion library that expresses the host genes and Gal4 DNA binding domain fusion bait vectors that express B m DNV ‐ Z genes. Y 2 H assay revealed 15 potential interactions between host and viral proteins. To verify the interactions, we modified and reconstructed a pair of bimolecular fluorescence complementation ( B i FC ) vectors and achieved the co‐expressions of the candidate host genes and viral genes in insect culture cells. The B i FC assay confirmed the specificity of the interactions including B. mori 35 kDa protease and two B m DNV ‐ Z proteins encoded by VD 1‐ ORF 2 and VD 2‐ ORF 1; B . mori transgelin and B m DNV ‐ Z protein encoded by VD 2‐ ORF 3; and B . mori serine protease precursor and B m DNV ‐ Z encoded by VD 2‐ ORF 3 in vitro . Our findings revealed that the specific host midgut proteins are involved in the interactions between B . mori and B m DNV ‐ Z , which will facilitate our understanding of the molecular mechanisms of B m DNV ‐ Z infection. Structured digital abstract BmDNV VD1-ORF4 physically interacts with trypsin-like protease by two hybrid pooling approach ( View interaction ) 35 kDa protease and BmDNV VD2-ORF1 physically interact by bimolecular fluorescence complementation ( View interaction ) 35 kDa protease and BmDNV VD1-...