Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
Fish Shellfish Immunol. 2013 Sep;35(3):957-64. doi: 10.1016/j.fsi.2013.07.009. Epub 2013 Jul 16.
White spot syndrome (WSS) is a viral disease caused by white spot syndrome virus (WSSV) which leads to severe mortality in cultured penaeid shrimp. In response to WSSV infection in Penaeus monodon, a Kazal serine proteinase inhibitor SPIPm2, normally stored in the granules of granular and semi-granular hemocytes is up-regulated and found to deter the viral replication. By using yeast two-hybrid screening, we have identified a viral target protein, namely WSV477. Instead of being a proteinase, the WSV477 was reported to be a Cys2/Cys2-type zinc finger regulatory protein having ATP/GTP-binding activity. In vitro pull down assay confirmed the protein-protein interaction between rSPIPm2 and rWSV477. Confocal laser scanning microscopy demonstrated that the SPIPm2 and WSV477 were co-localized in the cytoplasm of shrimp hemocytes. Using RNA interference, the silencing of WSV477 resulted in down-regulated of viral late gene VP28, the same result obtained with SPIPm2. In this instance, the SPIPm2 does not function as proteinase inhibitor but inhibit the regulatory function of WSV477.
白斑综合征（WSS）是由白斑综合征病毒（WSSV）引起的病毒性疾病，可导致养殖对虾严重死亡。在斑节对虾对 WSSV 感染的反应中，一种原本储存在颗粒状和半颗粒状血细胞颗粒中的丝氨酸蛋白酶抑制剂 SPIPm2 被上调，并被发现可阻止病毒复制。通过酵母双杂交筛选，我们鉴定出一种病毒靶蛋白，即 WSV477。与蛋白酶不同，WSV477 被报道为具有 ATP/GTP 结合活性的 Cys2/Cys2 型锌指调节蛋白。体外下拉测定证实了 rSPIPm2 和 rWSV477 之间的蛋白-蛋白相互作用。共聚焦激光扫描显微镜显示 SPIPm2 和 WSV477 在虾血细胞的细胞质中共定位。通过 RNA 干扰，WSV477 的沉默导致病毒晚期基因 VP28 的下调，与 SPIPm2 获得的相同结果。在这种情况下，SPIPm2 不作为蛋白酶抑制剂发挥作用，而是抑制 WSV477 的调节功能。
