The Regulation Mechanism Of E3 Ubiquitin Ligase HUWE1 On Zika Virus PrM Protein

Background: At present,Zika virus(ZIKV)is ubiquitous in the world.The epidemic situation has continued to develop,showing a trend of spreading and cross-border transmission.Vaccine development,clinical diagnosis and basic research for Zika virus disease are constantly advancing.ZIKV prM(precursor membrane)protein is the structural protein of ZIKV,which has an important impact on the structure,maturation process and pathogenicity of the virus.Objective: We hope to provide new ideas for the prevention and control of ZIKV infection and pathogenesis.Methods: In this study,the mechanism of ubiquitination and degradation of ZIKV prM protein by E3 ligase HUWE1 was explored by co-immunoprecipitation,immunoblotting and other techniques.Results: 1.ZIKV prM protein is similar in structure to DENV prM protein and both can be degraded in vivo;2.JEV prM protein is most closely related to WNV prM protein;3.ZIKV prM protein can be ubiquitinated and passed through the ubiquitin-proteasome pathway Degradation;4.ZIKV prM protein interacts with E3 ligase HUWE1;5.Gradient overexpression of HUWE1 promotes the degradation of ZIKV prM protein;6.Overexpression of HUWE1 accelerates the degradation of ZIKV prM protein and increases ubiquitin chains;7.Knockdown of HUWE1 makes ZIKV prM protein degradation is slowed down and ubiquitin chains are reduced;8.ZIKV prM protein is not degraded in SKN-SH cell line lacking HUWE1;9.ZIKV prM protein ubiquitination occurs in the prprotein region;10.ZIKV prM protein The 33 rd and 84 th lysine single mutants were more stable than the 85 th position;11.ZIKV prM protein lysine deletion double-site mutants were stable and attenuated ubiquitination compared with the wild type.Conclusion: 1.ZIKV prM protein is degraded by ubiquitinproteasome pathway;2.E3 ligase HUWE1 regulates the ubiquitination and degradation of ZIKV prM protein;3.ZIKV prM protein ubiquitination occurs at positions 33 and 84 on lysine residues.