Dechklar Manasave, Udomkit Apinunt, Panyim Sakol
Institute of Molecular Biology and Genetics, Mahidol University, 25/25 Phuttamonthon 4 Road, Salaya Campus, Nakhon Pathom 73170, Thailand.
Biochem Biophys Res Commun. 2008 Mar 21;367(4):768-74. doi: 10.1016/j.bbrc.2008.01.031. Epub 2008 Jan 15.
RNA interference (RNAi) has recently become a promising strategy for therapeutic of several viral diseases including those in the black tiger shrimp Penaeus monodon. However, the protein components that play role in RNAi in P. monodon have not yet been identified. Here, we report the cloning and functional characterization of a cDNA encoding Argonaute, a principal constituent of RNAi pathway in P. monodon. P. monodon's Argonaute (Pem-AGO) exhibited the two signature domains, PAZ and PIWI. Substantial level of Pem-ago expression could be suppressed by double-stranded RNA (dsRNA) that targeted PAZ coding sequence in shrimp primary culture of Oka cells. The Pem-ago depleted cells showed impaired RNAi as the expression of an endogenous gene was rescued from the dsRNA-mediated silencing in these cells. Our results imply that Pem-ago is required for effective RNAi in P. monodon and thus identify the first protein constituent of RNAi machinery in penaeid shrimp.
RNA干扰（RNAi）最近已成为治疗包括黑虎虾（斑节对虾）在内的几种病毒性疾病的一种有前景的策略。然而，在斑节对虾RNAi过程中发挥作用的蛋白质成分尚未被鉴定出来。在此，我们报道了斑节对虾RNAi途径的主要成分——一种编码AGO蛋白的cDNA的克隆及功能特性。斑节对虾的AGO蛋白（Pem-AGO）具有两个标志性结构域，即PAZ和PIWI。在斑节对虾Oka细胞原代培养物中，靶向PAZ编码序列的双链RNA（dsRNA）可显著抑制Pem-ago的表达。Pem-ago缺失的细胞表现出RNA干扰受损，因为这些细胞中内源性基因的表达从dsRNA介导的沉默中得以恢复。我们的结果表明，Pem-ago是斑节对虾有效RNA干扰所必需的，从而鉴定出了对虾RNA干扰机制中的首个蛋白质成分。
