Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Klong Luang, Pathumthani, Thailand.
Fish Shellfish Immunol. 2018 May;76:174-182. doi: 10.1016/j.fsi.2018.01.043. Epub 2018 Mar 1.
Hemocyte homeostasis-associated protein (PmHHAP) was first identified as a viral-responsive gene, due to a high upregulation in transcription following white spot syndrome virus (WSSV) infection. Functional studies using RNA interference have suggested that PmHHAP is involved in hemocyte homeostasis by controlling apoptosis during WSSV infection. In this study, the role of PmHHAP in host-viral interactions was further investigated. Yeast two-hybrid assay and co-immunoprecipitation revealed that PmHHAP binds to an anti-apoptosis protein, WSSV134. The viral protein WSSV134 is a late protein of WSSV, expressed 24 h post infection (hpi). Gene silencing of WSSV134 in WSSV-infected shrimp resulted in a reduction of the expression level of the viral replication marker genes VP28, wsv477, and ie-1, which suggests that WSSV134 is likely involved in viral propagation. However, co-silencing of PmHHAP and WSSV134 counteracted the effects on WSSV infection, which implies the importance of the host-pathogen interaction between PmHHAP and WSSV134 in WSSV infection. In addition, caspase 3/7 activity was noticeably induced in the PmHHAP and WSSV134 co-silenced shrimp upon WSSV infection. Moreover, PmHHAP and WSSV134 inhibited caspase-induced activation of PmCasp in vitro in a non-competitive manner. Taken together, these results suggest that PmHHAP and WSSV134 play a role in the host-pathogen interaction and work concordantly to control apoptosis in WSSV infection.
血淋巴细胞稳态相关蛋白（PmHHAP）最初被鉴定为一种对病毒有反应的基因，因为在感染白斑综合征病毒（WSSV）后，其转录水平显著上调。使用 RNA 干扰的功能研究表明，PmHHAP 通过控制 WSSV 感染期间的细胞凋亡参与血淋巴细胞稳态。在这项研究中，进一步研究了 PmHHAP 在宿主-病毒相互作用中的作用。酵母双杂交测定和免疫共沉淀表明，PmHHAP 与一种抗凋亡蛋白 WSSV134 结合。病毒蛋白 WSSV134 是 WSSV 的晚期蛋白，在感染后 24 小时（hpi）表达。在感染 WSSV 的虾中沉默 WSSV134 基因导致病毒复制标记基因 VP28、wsv477 和 ie-1 的表达水平降低，这表明 WSSV134 可能参与病毒的增殖。然而，PmHHAP 和 WSSV134 的共沉默抵消了对 WSSV 感染的影响，这意味着 PmHHAP 和 WSSV134 之间在宿主-病原体相互作用在 WSSV 感染中很重要。此外，在 WSSV 感染时，PmHHAP 和 WSSV134 共沉默的虾中 caspase 3/7 活性明显升高。此外，PmHHAP 和 WSSV134 以非竞争性方式体外抑制 caspase 诱导的 PmCasp 的激活。总之，这些结果表明，PmHHAP 和 WSSV134 在宿主-病原体相互作用中发挥作用，并协同作用控制 WSSV 感染中的细胞凋亡。
