RNF122 targets STING for ubiquitination at residues K95, K117, and K155 to regulate antiviral responses in a teleost fish

Ring finger protein 122 (RNF122), an E3 ubiquitin ligase, orchestrates antiviral immune responses in mammals by targeting retinoic acid-inducible gene 1 and melanoma differentiation-associated gene 5 for ubiquitination. However, its functional relevance in teleosts has yet to be clearly defined, particularly regarding the identification of substrate-specific regulatory sites. This study characterized RNF122 from mandarin fish ( <i>Siniperca chuatsi</i>), termed <i>sc</i>RNF122, and investigated its regulatory impact on stimulator of interferon genes (STING)-mediated antiviral signaling. Results showed that <i>sc</i>RNF122 expression was up-regulated in response to mandarin fish ranavirus (MRV) infection, and its overexpression suppressed <i>sc</i>STING-mediated interferon (IFN) production and enhanced MRV replication. Co-immunoprecipitation confirmed a direct interaction between <i>sc</i>RNF122 and <i>sc</i>STING. Functional assays demonstrated that <i>sc</i>RNF122 facilitated <i>sc</i>STING degradation through the ubiquitin-proteasome pathway, a process impeded by MG132 treatment. Ubiquitination analyses of various <i>sc</i>STING mutants revealed that <i>sc</i>RNF122 catalyzed <i>sc</i>STING ubiquitination at K95, K117, and K155 residues. Moreover, <i>sc</i>RNF122 significantly impaired <i>sc</i>STING-dependent antiviral responses by engaging negative regulatory elements within the signaling cascade. Overall, <i>sc</i>RNF122 was identified as a negative modulator of STING-mediated IFN signaling in mandarin fish, diminishing STING-dependent antiviral activity and promoting its degradation via the ubiquitin-proteasome pathway at lysine residues K95, K117, and K155. These findings provide mechanistic insight into the post-translational control of STING in teleosts and establish a foundation for future investigations into antiviral immune regulation.