| Encephalomyocarditis virus(EMCV)is a positive single-stranded RNA virus without envelope.Structural proteins VP1,VP2,VP3 and VP4 comprise the viral capsid,.Neutralizing antigens locate mainly in VP1,VP2 and VP3,among them,VP1 has the strongest antigenicity.At present,the area of EMCV is more and more popular all over the world,and infection rate is increasing year by year.However,the mechanism of infection and host range limitation remain unclear.In this study,we screened HuvEc cellular proteins interacting with VP 1,VP2 and VP3 respectively by yeast two hybrid.Further we analyzed the subcellular localization,molecular functions and biological processes of the interactors.In this study,we screened HuvEc cDNA library with bait VP1,VP2,and VP3 respectively with three replicates by using split-ubiquitin yeast two hybrid system.After yeast β-galactosidase assays and yeast transforming tests,false positive clones were removed,and finally we obtained 57 interacting proteins.There are 15 proteins that interact with VP1,and there are 39 interacting proteins with VP2,and VP3 has an interaction protein of about 3.We found out that BNIP3、C14orf1、SERP1、UBC、YIPF6 interacted with both VPI and VP2,UBB interacted with both VP1 and VP3,SPCS1、ZUFSP interacted with all the three capsid proteins.The interacting proteins were mainly distributed in the cytoplasm,cell membrane,nucleus,mitochondria,Golgi,endoplasmic reticulum and other organelles.The molecular function of these proteins were including binding,catalytic activity,channel regulator activity,structural molecule activity,translation regulator activity,and transporter activity.The major biological processes involved were biological control,cell formation,cell metabolism,cell proliferation,cell localization,transport and immune response.The preliminary screening of this study to provide a reference for studying the molecular mechanism of EMCV infection and viral pathogenesis. |
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