Study On The Mechanism Of SABHD16A Regulating IFN Against JEV

Interferon(IFN)signaling pathway is an important component of innate immune response.After interferon is induced,it will up-regulate the expression of a series of interferon-stimulated genes(ISGs).Interferon-induced transmembrane proteins(IFITMs)are a class of small molecule membrane proteins.Studies have shown that IFITMs play an important role in innate antiviral immune response and are the first line of defense against the invasion of a variety of viruses.Japanese encephalitis(JE)is a zoonosis caused by Japanese encephalitis virus(JEV).Swine are intermediate hosts of JEV.Our previous study proved that swine IFITM protein has against JEV effect depends on palmitoylation modification Swineα/β hydrolase domain containing 16A(sABHD16A)interacts with s IFITMs and regulates the palmitoylation cycle of s IFITMs.But the molecules that regulate sABHD16 A have not been identified.Lehner et al.used a high-throughput yeast two-hybrid system to find that human ABHD16 A interacts with RING-finger protein5(RNF5).However,whether this interaction exists in swine and whether sRNF5 regulates the function of sABHD16 A through ubiquitination has not been reported.Therefore,exploring the regulation mode of sRNF5 on sABHD16 A will help to better understand the mechanism of sABHD16 A regulating IFN against JEV.In this study,with the help of novel palmitoylation detection technology of APEGS,protein interaction analysis method of Bi FC and Co-IP and other methods,we found that:1.sABHD16 A reduces the against JEV ability of s IFITM1 by depalmitoylating s IFITM1;2.sABHD16 A promotes the production of Lyso-PS in cells,and the specific inhibitor KC01 can reduce the level of s IFITM1 depalmitoylation and negatively regulate the against JEV ability of s IFITM1.3.sRNF5 and sABHD16 A interact with each other and show intracellular colocalization.4.sRNF5 affects the stability of sABHD16 A by ubiquitination,which weakens the ability of sABHD16 A to negatively regulate JEV infection.The analysis of s IFITM1-sABHD16A-sRNF5 interaction in this study helps to understand the regulation of sABHD16 A in IFN against JEV mechanism from a new perspective,and provides a theoretical basis for the study and introduction of novel JE prevention and intervention measures.