Journal
VIROLOGY JOURNAL
Volume 20, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s12985-023-02026-x
Keywords
Cyprinid herpesvirus 2; ORF55; Prokaryotic expression; Phage display technology; Thymidine kinase; Virulence factor
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This study investigated the biological function of the open reading frame 55 (ORF55) gene of Cyprinid herpesvirus 2 (CyHV-2) in viral replication. The purified CyHV-2 ORF55 protein was obtained using prokaryotic expression, and interacting peptides were identified through phage display. Host interacting proteins were predicted and validated.
Background Cyprinid herpesvirus 2 (CyHV-2) is a pathogenic fish virus belonging to family Alloherpesviridae. The CyHV-2 gene encoding thymidine kinase (TK) is an important virulence-associated factor. Therefore, we aimed to investigate the biological function of open reading frame 55 (ORF55) in viral replication. Methods Purified CyHV-2 ORF55 protein was obtained by prokaryotic expression, and the interacting peptide was screened out using phage display. Host interacting proteins were then predicted and validated. Results ORF55 was efficiently expressed in the prokaryotic expression system. Protein and peptide interaction prediction and dot-blot overlay assay confirmed that peptides identified by phage display could interact with the ORF55 protein. Comparing the peptides to the National Center for Biotechnology Information database revealed four potential interacting proteins. Reverse transcription quantitative PCR results demonstrated high expression of an actin-binding Rho-activating protein in the latter stages of virus-infected cells, and molecular docking, cell transfection and coimmunoprecipitation experiments confirmed that it interacted with the ORF55 protein. Conclusion During viral infection, the ORF55 protein exerts its biological function through interactions with host proteins. The specific mechanisms remain to be further explored.
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