The Role Of Viral Induced SNX10 In Regulation Of Influenza Virus Infection

BackgroundSeasonal influenza and influenza pandemics are serious threats to public health and the global economy.Current drugs against influenza are mainly targeted to two viral proteins,M2 ion channel blockers(such as amantadine)and neuraminidase inhibitors(such as oseltamivir),but some new influenza virus strains have become resistant to these drugs.There is an urgent need to develop antiviral targets.Since influenza virus infection requires the assistance of a large number of host factors,host factors critical to virus replication become potential drug targets.The entry of influenza virus into the host cell is the first step of infection.Inhibition of influenza virus entry can effectively block virus infection.In addition to targeting the necessary viral proteins(such as the hemagglutinin protein HA),it can also target host proteins which affect virus entry.As previously reported,SNX10 is mainly distributed on the membrane of endosome.Interacted with-v-ATPase,SNX10 can activate the proton pump to induce acidic vacuoles.Furthermore,v-ATPase is the intracellular major proton pump,and it induced acidic endosome is crucial for virus entry.The acidic vacuoles regulated by SNX10 is similar to the acidic endosome induced by influenza viras.Therefore,we hypothesize that SNX10 may induce acidic endosome by binding to v-ATPase after virus infection and then promotes virus enter into the cells.More importantly,due to the similar pathways in many other virases entry,inhibition of SNX10 may lead to the development of broad-spectrum antiviral drugsObjectives:In this study,SNX10 as the research object,we explored the role of SNX10 in regulating IAV infection,and further elucidated its mechanism.In this project,we are committed to discover host factors that against influenza virus and aim to provide evidence and ideas for finding new antiviral targets.Methods:1.RT-PCR,Western blotting,CHX and MG-132 treatment,ubiquitination assay were used to explore the expression of SNX10 after IAV infection.2.RNAi assay,overexpression assay,plaque assay and indirect immunofluorescence assay were devised to detect the regulation of SNX10 in IAV infection.3.Neutral red,Lysotracker red,confocal,immunofluorescence colocalization experiments were utilized to study the mechanism of SNX10 in regulating IAV infection.Results:1.IAV infection or Poly(I:C)stimulation up-regulated SNX10 expression in cells.After IAV infection,there was no significant change in the expression of SNX10 mRNA,but the ubiquitination of SNX10 was downregulated.2.After knockdown SNX10,IVA NP expression in cells was significantly reduced.In contrast,IVANP expression was increased after overexpression of SNX10.3.Depletion of SNX10 resulted in a reduction of NP expression in the nuclei.However,overexpression of SNX10 promoted NP expression in the nuclei.4.After overexpression of SNX10,vacuoles are formed in cells,and the interaction of SNX10 with v-ATPase induced acidic vacuoles.5.IAV infection induced acidic endosome in cells.SNX10 co-localized with the acidic endosome induced by IAV,and knockdown of SNX10 could inhibit the production of acidic endosome after IAV infection.Conclusion:1.IAV infection could stabilize the expression of SNX10 in cells by inhibiting the ubiquitination degradation of SNX10.2.SNX10 regulated the infection of influenza viruses.3.SNX10 affected the early step during influenza virus infection.4.SNX10 induced acidic vacuoles in cells.5.SNX10 was involved in the acidic endosome required for virus entry.