Tumor Susceptibility Gene 101 Functions In The Process Of WSSV Infection Of Marsupenaeus Japonicus

The cellular endosomal sorting complexes required for transport(ESCRTs),comprised of five distinct components(ESCRT-0,-?,-?,-? and Vps4)involve in many different physiological processes.ESCRT-I protein,the tumor susceptibility gene 101(TSG101)is connected with entry,transport and release of many different kinds of virus.However,whether TSG101 participates in the infection of White Spot Syndrome Virus(WSSV)remains unknown.In this study,we firstly identified TSG101 from kuruma shrimp,Marsupenaeus japonicus,designated AjTSG101,which was drastically upregulated both at mRNA and protein levels after WSSV infection.Knockdown of MjTSG101 expression by RNA interference inhibited the virus replication,whereas "overexpression" of MjTSG101 increased WSSV replication in the shrimp.That is,WSSV is able to utilize MjTSG101 related machinery to facilitate its replication.Meanwhile,several components that are capable of interacting with TSG101(Vps28,Hrs and Alix)of ESCRTs were significantly upregulated post WSSV infection,which suggested that these molecules involve in the infection of WSSV together with TSG101.Immunocytochemistry analysis found that MjTSG101 and WSSV were colocalized in hemocytes.Expression level of Rab5 and Rab7(marker of early and late endosomes respectly)decreased significantly after silencing of MjTSG101.At the same time,knockdown of Rab5 and Rab7 expression by RNA interference diminished the virus replication.All the results suggested that MjTSG101 participates in the transportation of WSSV through early and late endosomes pathway.Subsequently,observation of MjTSG101's subcellular location found that MjTSG101 gathered around plasma membrane at 24 h post WSSV infection,implied that MjTSG101 plays a role at late stage of WSSV infection.Addition of N16(an inhibitor of HIV-1 assmbly and budding)remarkably decreased the replication of WSSV.All these indicated that WSSV is able to hijack MjTSG101 associated with some other ESCRT components to complete its release.