Identification and antiviral mechanism of a novel chicken-derived interferon-related antiviral protein targeting PRDX1

In this study, we identified a new chicken-specific protein, named chicken interferon-related antiviral protein (chIRAP) after sequence analysis and comparison, which inhibited the proliferation of various viruses including influenza A virus (IAV) and Newcastle Disease Virus (NDV) in vitro, and chicken embryos with high expression of chIRAP reduced IAV infection. Mass spectrometry analysis of chIRAP interacting proteins and screening of interacting proteins affecting the function of chIRAP revealed that the deletion of endogenous chicken peroxiredoxin 1 (chPRDX1) significantly reduced the antiviral effect of chIRAP. In order to clarify the functional site of chPRDX1 affecting the antiviral effect of chIRAP, we constructed the point mutants of chPRDX1 based on the results of molecular docking (D79A, T90A, K93A, Q94A, R110A, R123A), and screened the sites affecting the antiviral effects of chIRAP by knockdown of endogenous chPRDX1 combined with the overexpression mutant strategy, the results showed that the mutations in the sites affected the antiviral effects of chIRAP to different degrees, with D79A being the most significant, and the D79A mutation of chPRDX1 reduces the ability of chPRDX1 to regulate reactive oxygen species (ROS). chIRAP may exert antiviral effects by regulating the intracellular ROS balance at the D79 site of chPRDX1. In conclusion, we identified a novel chicken-derived antiviral protein, clarified its antiviral effects and preliminarily explored its mechanism of action, which provides a new tool and option for the prevention and treatment of avian-origin viral diseases, especially avian-origin related zoonotic diseases.