Screening of Goose Parvovirus Vp1 and Interacting Proteins in Infected Host Cells and Correlation with Goose Parvovirus Replication

The VP1 gene of goose parvovirus regulates the release of viral capsid proteins and daughter viruses. Screening host cells for proteins that interact with viral VP1 and probing the regulatory mechanisms of goose parvovirus replication provides a basis for outbreak control of goose parvovirus. In this study, a high-quality yeast two-hybrid (Y2H) cDNA library was constructed and validated using duck embryonic fibroblasts in order to screen for interaction partners of goose parvovirus VP1 protein in duck embryonic fibroblasts (DEFs). Self-activation of VP1 was detected by phenotypic screening, and positive clones were obtained by Y2H screening. Transformation experiments further validated this. Blast was used to analyze the sequences of the positive clones. Eukaryotic expression vectors were constructed after cloning eEF1A from the analyzed sequences. The interaction between VP1 and eEF1A was verified by immunoprecipitation and immunofluorescence. The association of eEF1A with goose parvovirus replication was investigated by Real-time PCR, viral titer assay and Trypan blue staining. The results showed an interaction of eEF1A with VP1, and overexpression of eEF1A could promote the replication of goose parvovirus, and silencing of eEF1A could inhibit the replication of goose parvovirus. The results of this study provide a basis for further study of the replication mechanism of goose parvovirus. The presence of eEF1A interacting with the VP1 gene in GEFs and the presence of eEF1a is importantly associated with GPV proliferation.