Study On The Function Of Host Cell Adaptor Protein Complex In Rabies Virus Infection

Rabies is an infectious disease which is extremely dangerous all over the world.More than 60,000 people die of rabies every year in the world and the pathogenesis of RABV has not fully clarified.The pathogen of rabies is RABV(Rabies virus,RABV),and RABV belongs to the capsule virus of the Rhabdoviridae family(Lyssaviruses).It encodes five structural proteins,nuclear protein(N),large protein(L)matrix protein(M),glycoprotein(G)and phosphorylated protein(P).G protein is one of the most important structural proteins of RABV.It plays an important role in the early stage of RABV infection.It can bind to the receptors on the cell membrane and mediate the entry of RABV into cells.AP:(Adaptor Protein Complex):The adaptor related protein complex is a polymer whose main physiological function is to separate the cargo proteins into the specific membrane intervals of the cell.At present,four kinds of adaptor related protein complex have been found in mammalian cells.AP2(adaptor complex 2)is the most important adapter protein complex in cells.It can participate in the clathrin mediated endocytosis and link the cargo protein(cargo)and clathrin.The previous study in our laboratory had screened host genes related to the virus life cycle of RABV in the human genome using RNA interference(RNAi).The RAVB infection rate was decreased when AP2M1 gene was knockdown.To explore the mechanism that AP2M1 affected RABV infection.When AP2M1 gene was knocked-down or over-expressed,the infection rate of rabies virus in HEK-293 cells were analyzed by high intension screening and virus titer detection.We proved that AP2M1 gene played a role in the early stage of RABV infection by acid bypass experiments.The interaction between AP2M1 and RABV G protein was investigated by co-immunoprecipitation experiments and laser scanning confocal microscopy.The results showed that The RAVB infection rate was decreased when AP2M1 gene was knockdown.We found that AP2M1 protein was not co-located with G protein.The co-immunoprecipitation experiments illustrated that there was no interaction between G protein and AP2M1 protein.To sum up,the AP2M1 gene played a role in the early stage of RABV infection.However,there was no interaction between G protein and AP2M1 protein.This study provides a theoretical basis to find the specific role of AP2M1 in the RABV life cycle.Vesicular Stomatitis virus(VSV)is a member of Rhabdoviridae.The VSV infection rate was decreased when AP2M1 gene was knocked-down and over-expressing of AP2M1 could increase the infection rate of VSV.It is suggested that the AP2M1 gene can affect VSV infection.This study provides a theoretical basis to find a new potential drug target for virus.