Preliminary Study On The Role Of Host Cell EEF1A In Goose Parvovirus Infection

Gosling plague was an acute,subacute septic epidemic of goslings.It mainly infected goslings and young muscovy ducks of 3-20 days old.It spreaded rapidly and had a high mortality rate.The disease was not only popular in China all the year round,but also caused serious losses.Goose parvovirus(GPV)was the pathogen of gosling plague(GP).The GPV genome was about 5.2 kb in length and was single-stranded,linear DNA.The left side encoded the non-structural proteins NS1 and NS2,which were mainly involved in viral replication and transcriptional regulation;the right side encoded capsid proteins VP1,VP2 and VP3,of which VP1 included the entire gene sequence of VP2 and VP3.The translation elongation factor1A(eEF1A)was a protein with a variety of biological functions and was abundant in cells,second only to actin content,and its genes and expressions were strictly conserved in different species.eEF1A was not only a protein that translation must participate in,but more importantly,it was involved in many key cell growth processes and diseases,such as translational control,apoptosis,signal transduction,viral replication,oncogene transformation and cytoskeletal composition and so on.GPV VP1 protein played an important role in the process of virus adsorption and invasion,but there has been little research on the interaction between VP1 protein and host cells so far.In this study,the eukaryotic expression vectors pcDNA3.0-VP1 and pcDNA3.1-eEF1A were cloned by cloning the VP1 gene and the eEF1A gene,and then the interaction between VP1 and eEF1A was verified.The interaction between VP1 and eEF1A was confirmed by immunoprecipitation assay and immunofluorescence assay.The effect of host cell eEF1A on GPV replication was investigated by Real-time PCR,virus titer assay and trypan blue staining assay.Firstly,eEF1A was transfected into goose embryo fibroblasts to establish a cell system that overexpresses eEF1A,the results showed that viral replication increased after overexpression of eEF1A in 24 h,48h and 72 h,Subsequently,we designed and synthesized an interfering siRNA molecule targeting eEF1A protein,and down-regulated the expression of eEF1A by siRNA in 24 h,48h and 72 h.The results showed that silencing eEF1A protein inhibited the increase of GPV and genome replication.In conclusion,this study identified the interaction between GPV VP1 protein and the eEF1A protein,GPV replication was promoted by overexpressing eEF1A and GPV was suppressed by silent expression of eEF1A.