Screening and identification of cyprinid herpesvirus 2 (CyHV-2) ORF55-interacting proteins by phage display

• Published in: Virology journal 2023-04, Vol.20 (1), p.66-66, Article 66
• Main Authors: Qian, Min, Xiao, Simin, Yang, Yapeng, Yu, Fei, Wen, Jinxuan, Lu, Liqun, Wang, Hao
• Format: Article
• Language: English
• Subjects: Actin; Analysis; Animals; Antibodies; bacteriophages; Bacteriophages - genetics; Biotechnology; Carps; Case Report; Cloning; Cyprinid herpesvirus 2; E coli; family; Fish Diseases; Gene amplification; genes; Herpes viruses; Herpesviridae - genetics; Herpesviridae Infections; Herpesviruses; Identification and classification; Infections; Kinases; Medical research; Molecular Docking Simulation; National Center for Biotechnology Information; Open Reading Frames; ORF55; Peptides; Phage display; Phage display technology; Polypeptides; precipitin tests; prediction; Prokaryotic expression; Proteins; reverse transcriptase polymerase chain reaction; Reverse transcription; Thermal cycling; Thymidine; Thymidine kinase; Transfection; vertebrate viruses; Viral infections; Viral proteins; Virulence; Virulence (Microbiology); Virulence factor; virus replication
• ISSN: 1743-422X; 1743-422X
• DOI: 10.1186/s12985-023-02026-x

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. 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. 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. During viral infection, the ORF55 protein exerts its biological function through interactions with host proteins. The specific mechanisms remain to be further explored.