The Role Of Ribosomal L12 And S20 Proteins In The Replication Of Porcine Reproductive And Respiratory Syndrome Virus

Reproductive and respiratory syndrome(Porcine reproductive and respiratory syndrome,PRRS),also known as blue-ear disease,quickly spread to almost all pig-raising countries after it was first reported in 1986,causing huge economic losses to the global pig industry.Porcine Reproductive and Respiratory Syndrome Virus(PRRSV)is an enveloped virus.The envelope protein plays an important role in the process of virus infection,assembly and release.Among them,PRRSV GP2 b is a glycosylated membrane protein that participates in virus invasion and replication,and is highly conserved and relatively abundant in the two types of PRRS virus particles.Therefore,GP2 b may become a genetic target for the prevention and control of PRRSV.One point,given that the GP2 b protein plays an important role in the replication cycle of the porcine reproductive and respiratory syndrome virus,our laboratory has screened the host ribosomal proteins L12 and S20 that interact with GP2 b through yeast two-hybrid in the early stage,and they are involved in the replication of PRRSV.The role of is not yet clear.In this study,immunoprecipitation and laser confocal tests were used to test whether ribosomal protein interacts with PRRSV GP2 b protein in cells.The effects of PRRSV on the expression of ribosomal protein after infection of cells with PRRSV were tested by fluorescence quantitative PCR and Western blotting.,And the effects of overexpression and knockdown of RPL12 and RPS20 on the replication of PRRSV in Marc-145 cells,and construct RPL12 and RPS20 truncated plasmids to verify their functional regions.The resultshowed that there is an interaction between ribosomal protein L12 and PRRSVGP2 b protein,and they are co-localized in the cytoplasm.PRRSV can up-regulate the expression of RPL12 in cells after infection.With the replication of PRRSV,the transcription level and protein level of RPL12 increased significantly.Further research found that overexpression of RPL12 can promote the replication of PRRSV,while knockdown of RPL12 can inhibit the replication of PRRSV.The RPL12 truncated plasmid was constructed and found that its pro-viral region was located between 66-166 aa.There is an interaction between the ribosomal protein S20 and the GP2 b protein,and they co-localize in the cytoplasm.PRRSV infection can up-regulate the expression of RPS20.With the replication of RRSV,the transcription level and protein level of RPS20 are significantly enhanced.Overexpression of RPS20 promotes the replication of PRRSV in Marc-145 cells.Knockdown of RPS20 can inhibit the replication of the virus.A truncated RPS20 plasmid was constructed and the virus-promoting region was found to be between 40-80 aa.This study found for the first time that PRRSV infection can up-regulate the expression of RPL12 and RPS20 in cells,which interact with PRRSV GP2 b protein and co-localize in the cytoplasm.Overexpression of RPL12 can promote the replication of PRRSV,and the virus-promoting region is between 66-166 aa,and overexpression of RPS20 can promote the replication of PRRSV,and the virus-promoting region is between 40-80 aa.These results are for further research on the infection and replication of PRRSV.And the interaction with the host provides a new entry point,which provides a useful exploration for the research of antiviral drugs.