ZNF205 Positively Regulates RLR Antiviralsignaling By Targeting RIG-Ⅰ

Pattern Recognition Receptors(PRRs)are able to directly recognize pathogens and,by recognizing and binding to ligands,provide natural immune protection against infections,tumors and other diseases.Different PRRs recognize different ligands,and RIG-Ⅰ like receptors(RIG-Ⅰ like receptors)are an important cytoplasmic RNA recognition receptor.Among them,RIG-Ⅰ recognizes viral RNA and interacts with the adapter protein VISA,which subsequently activates downstream signaling and induces type I interferon and pro-inflammatory cytokine production.The activity of RIG-Ⅰ,a key to signal initiation,is tightly regulated to avoid excessive and uncoordinated cytokine production.To further explore the regulatory mechanism of RIG-Ⅰ,we screened and identified ZNF205 as a novel interacting protein of RIG-Ⅰ using yeast two-hybrid technique.Through Dual-luciferase reporter assay,we found that overexpression of ZNF205 promotes Se V-induced activation of the IFN-β-promoter and ISRE.And at the m RNA level,overexpression of ZNF205 could effectively enhance the transcription of downstream antiviral genes IFNBI and CXCL10.In addition,overexpression of ZNF205 suppressed the replication of VSV-GFP virus.After that,we knocked down ZNF205 using CRISPR Cas9 technology and further verified that knockdown of ZNF205 would inhibit type I interferon production and cellular antiviral ability by q PCR,native-PAGE,and plaque assay.Further experiments revealed that the zinc finger domain of ZNF205 interacted with the RIG-Ⅰ CARD domain and enhanced Se V-induced activation of IFN-β-promoter and ISRE.Mechanistic experiments showed that ZNF205 enhanced the polyubiquitination of the K63 linkage of the RIG-Ⅰ CARD domain and promoted the ubiquitination of RIG-Ⅰ by primarily targeting residues K164 and K172.In summary,this study reveals that ZNF205 positively regulates the antiviral natural immune response of RLR by promoting the ubiquitination modification of K164 and K172 sites on RIG-Ⅰ.Currently,there are not many studies on the function of ZNF205 in innate immunity.In the present study,we identified for the first time the positive regulatory role of ZNF205 in the RIG-Ⅰ-mediated antiviral signaling pathway.However,more in-depth exploration is needed about the fine mechanism of ZNF205 regulation of RIG-Ⅰ.