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Open Access 01-12-2023 | Herpes Virus | Case Report

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

Authors: Min Qian, Simin Xiao, Yapeng Yang, Fei Yu, Jinxuan Wen, Liqun Lu, Hao Wang

Published in: Virology Journal | Issue 1/2023

Abstract

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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Title: Screening and identification of cyprinid herpesvirus 2 (CyHV-2) ORF55-interacting proteins by phage display
Authors: Min Qian
Simin Xiao
Yapeng Yang
Fei Yu
Jinxuan Wen
Liqun Lu
Hao Wang
Publication date: 01-12-2023
Publisher: BioMed Central
Keyword: Herpes Virus
Published in: Virology Journal / Issue 1/2023
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/s12985-023-02026-x

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