白斑综合症病毒极早期蛋白WSV403相互作用蛋白的筛选

Screening for the interactors of WSSV immediate early protein WSV403

摘要: WSV403是白斑综合症病毒(WSSV)的一个极早期基因，它编码一种E3泛素连接酶。在293T细胞中，利用串联亲和纯化和蛋白质谱技术筛选WSV403的相互作用蛋白，获得了15个可能与WSV403结合的蛋白。在此基础上对5个候选基因进行了克隆，并分别将它们与WSV403基因共表达。通过免疫共沉淀和免疫荧光分析证实了WSSV极早期蛋白WSV403能够与核转运蛋白(KPNA6)、丝/苏氨酸蛋白磷酸酶2A 65 kDa调节亚基A(PPP2RIA)和striatin(STRN)发生相互作用。该研究结果为进一步揭示WSV403的作用机制奠定了基础。

Abstract: WSV403 is an immediate early gene of white spot syndrome virus (WSSV) which encodes an E3 ubiquitin ligase. We screened for the interactors of WSV403 in 293T cells using tandem affinity purification and protein spectrometry. Fifteen proteins that might bind to WSV403 were obtained. On this basis, we cloned 5 candidate genes and co-expressed them with WSV403. Co-immunoprecipitation and immunofluorescence assays confirmed that the immediate early protein WSV403 can interact with karyopherin subunit alpha 6 (KPNA6), serine/threonine protein phosphatase 2A 65 kDa regulatory subunit A (PPP2RIA), and striatin (STRN). The results provide directions for revealing the role of WSV403 in WSSV infection.