The Research Of Glycoprotein Of Spring Viraemia Of Carp Virus Interacted With 14-3-3β/α-A

Spring Viraemia of Carp Virus(SVCV)is a genus of Sprivivirus belonging to the family Rhabdoviridae.It is a contagious pathogen that endangers the safety of cyprinoid breeding.It can cause typical spring viraemia of carp(SCV)after infection with fish.The disease is a common epidemic in European,Asian,and North American cyprinoid breeding areas.Although the disease has not yet been reported in China as a large-scale breaking out disease case,it is one of the largest countries in the world for aquaculture.Disease poses a potentially huge threat to China’s aquaculture industry.SVCV infection causes fish body surface bleeding,ascites,and inflammation of the venthole,and severe cases until death.At present,the pathogenic mechanism and entry mechanism of SVCV are still unknown.Recent studies have shown that the interaction of the SVCV protein with the host protein has an important effect on the immune escape and replication of the virus.SVCV structural protein glycoprotein(Glycoprotein,Gp)located on the viral envelope mainly determines the serotype of the virus and plays a role in endocytosis of the virus when it invades cells.Therefore,studying the interaction between G protein and host protein of SVCV has extremely important scientific significance in revealing the entry and pathogenic mechanism of the virus,as well as the development of antiviral drugs and prevention vaccines.Firstly,using tandem affinity purification(TAP)combined with mass spectrometry(MS),FHM cells were screened for proteins that may interact with SVCV G protein.Secondly,Co-immunoprecipitation(Co-Immunoprecipitation,Co-IP)was used to verify the results of the experiments.The results showed that SVCV Gp can interact with 14-3-3β/α-A protein.Finally,14-3-3β/α-A was highly conserved among mammals,birds,amphibians and fishes by the alignment and homology analysis of the 14-3-3β/α-A protein sequence.One of the proteins has a homology of up to 97%.After FHM cells were infected with SVCV,the protein and m RNA levels of 14-3-3β/α-A were down-regulated.Overexpression of 14-3-3β/α-A in FHM cells enhances attachment and entry of SVCV,however,14-3-3β/α-A has no effect on SVCV replication during late infection.This article demonstrated for the first time that the interaction between 14-3-3β/α-A and SVCV G protein,and can promote the entry and attachment of SVCV,which provides a new horizon for revealing the pathogenic mechanism of SVCV and the development of antiviral drugs.