Jang Kyoung Jin, Lee Na-Ra, Yeo Woon-Seok, Jeong Yong-Joo, Kim Dong-Eun
Department of Biomaterial Control, Dong-Eui University, Busan 614-714, Republic of Korea.
Biochem Biophys Res Commun. 2008 Feb 15;366(3):738-44. doi: 10.1016/j.bbrc.2007.12.020. Epub 2007 Dec 17.
Recent outbreak of Severe Acute Respiratory Syndrome (SARS) that caused almost 800 victims requires a development of efficient inhibitor against SARS coronavirus (SCV). In this study, RNA aptamers against SCV NTPase/Helicase (nsP10) were isolated from RNA library containing random sequences of 40 nts using in vitro selection technique. Nucleotide sequences of enriched RNA aptamer pool (ES15 RNA) contain AG-rich conserved sequence of 10-11 nucleotides [AAAGGR(G)GAAG; R, purine base] and/or additional sequence of 5 nucleotides [GAAAG], which mainly reside at the loop region in all the predicted secondary structures. Isolated RNAs were observed to efficiently inhibit double-stranded DNA unwinding activity of the helicase by up to approximately 85% with an IC(50) value of 1.2nM but show a slight effect on ATPase activity of the protein in the presence of cofactor, poly (rU). These results suggest that the pool of selected aptamers might be potentially useful as anti-SCV agents.
近期爆发的严重急性呼吸综合征(SARS)导致近800人死亡,这需要开发一种针对SARS冠状病毒(SCV)的有效抑制剂。在本研究中,利用体外筛选技术从包含40个核苷酸随机序列的RNA文库中分离出针对SCV NTPase/解旋酶(nsP10)的RNA适配体。富集的RNA适配体库(ES15 RNA)的核苷酸序列包含10 - 11个核苷酸的富含AG的保守序列[AAAGGR(G)GAAG;R,嘌呤碱基]和/或5个核苷酸的附加序列[GAAAG],这些序列主要位于所有预测二级结构的环区。观察到分离出的RNA能有效抑制解旋酶的双链DNA解旋活性,抑制率高达约85%,IC(50)值为1.2nM,但在辅因子聚(rU)存在的情况下,对该蛋白的ATPase活性影响较小。这些结果表明,所选适配体库可能有潜力作为抗SCV药物。