| Encephalomyocarditis virus is a member of the Cardiovirus genus of the Picornaviridae family.It has a wide host spectrum,which can cause acute myocarditis,encephalitis,diabetes,reproductive disorders and other symptoms,causing huge economic losses to livestock breeding at home and abroad and also harm to public health.Although more and more studies have been done on EMCV in recent years,the infection mechanism and pathogenic mechanism of the virus are still unclear.This study is mainly based on the heat shock protein 90β,which interacts with virus protein screened by yeast two-hybrid system and virus overlay protein binding assay(VOPBA),in order to clarify the regulatory role and mechanism of heat shock protein 90β in encephalomyocarditis virus infection in vitro.EMCV was inoculated after up-regulating or down-regulating the expression of HSP90β in cells.The effect of HSP90 β on the proliferation of EMCV in vitro was determined by detecting virus copy number and virus titer.Western blot and q-PCR experiments were used to further explore the specific mechanism of the regulatory effect of HSP90β on EMCV.The findings are as follows:1.Up-regulation of host HSP90β expression significantly promoted the proliferation of EMCV in vitro.On the contrary,down-regulation of HSP90β by siRNA obviously inhibited the multiplication of EMCV,and GA,an inhibitor of HSP90β,also obtained the similar result as RNAi assay.2.EMCV-VP2 co-localized and interacted with HSP90β.Overexpression of VP2 promoted the expression of HSP90β and inhibited the production of IFN-β.3.Up-regulation of HSP90β inhibited the production of IFN-β,and down-regulation of HSP90β by siRNA promoted the production of IFN-β.HSP90β inhibited the production of IFN-β by inhibiting MAVS,TBK1 and IRF3 expression in a concentration dependent manner.Otherwise,HSP90β co-localized and interacted with MAVS,TBK1 and IRF3.These results clarified that after EMCV infection,HSP90β was hijacked by structural protein VP2,through up-regulating the expression of HSP90β to inhibit expression of MAVS,TBK1 and IRF3 in IFN-β signaling pathway.Then restrained IFN-β production to promote the proliferation of EMCV in vitro.This study revealed a new escape mechanism of EMCV infection,which laid a foundation for further development of relevant researches,and also provided a new sight for the prevention and control of encephalomyocarditis virus infection.In the future,we can try to use HSP90β inhibitors for anti-encephalomyocarditis virus infection related treatment. |
