Study On The Mechanism Of Host Factor KCNMA1 Affecting Influenza Virus Replication

Animal influenza has caused great economic losses to the global livestock industry,it can infect humans across host barriers,which cause serious public health problems,and the pathogen of the disease is the influenza virus.Influenza viruses constantly mutate during evolution and their genomic mutations and recombinations will cause antigen drift and antigen transformation,which makes the virus gain the ability to infect more new hosts and continue to produce new viruses that endanger animal and human health.On the account of this feature,the vaccines developed against the virus need to be updated as the virus evolves,and drugs which target viral proteins gradually become resistant.Therefore,it is particularly important to explore the mechanism of the interaction between host and influenza virus during the period against the virus.In this study,to evaluate the effects of replication of six influenza viruses(WSN,GD1330,FJ1660,AH1,SD008-627 K and SD007)including H1,H5,H7 and H9 subtypes,we use the combination of KCa1.1 small molecule antagonist Paxilline and siRNA interference method to perform functional inhibition and down-regulation of the host factor KCNMA1 on A549 and 293 T cell lines,respectively.The results show that KCa1.1 encoded by KCNMA1 can help the influenza virus establish infection on cells.In order to explore the specific stage of the host factor KCNMA1 affecting the life cycle of influenza virus,the endocytosis efficiency of influenza viruses H1,H5,H7,and H9 subtypes were evaluated in A549 cell lines,and the results showed that KCa1.1 can help all subtypes of influenza viruses complete the internalization process.Influenza virus HA protein plays a key role in the process of endocytosis of virions.In order to further explore the mechanism of KCa1.1 and related pathways,this study used Immunoprecipitation(IP)combined with shotgun mass spectrometry to screen and the host membrane proteins that interact with HA protein of H5 and H7 subtype influenza virus,and gene ontology(GO)analysis was performed on the screened protein database.The host membrane proteinS that interacts with KCa1.1 in the case of infection are also identified,which provides basic data for further revealing the mechanism of KCa1.1 helping the influenza virus to complete endocytosis.Relevant research on the early invasion of host cells by influenza viruses is not yet complete.This study reveals that the host protein KCa1.1 can affect the internalization process of various subtypes of influenza viruses,enriching the life cycle network of interaction between influenza viruses and hosts.The host factor KCNMA1 has potential as a new drug target.