Isolation of inhibitory RNA aptamers against severe acute respiratory syndrome (SARS) coronavirus NTPase/Helicase.
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.
Jang
Kyoung Jin
KJ
Department of Biomaterial Control, Dong-Eui University, Busan 614-714, Republic of Korea.
Lee
Na-Ra
NR
Yeo
Woon-Seok
WS
Jeong
Yong-Joo
YJ
Kim
Dong-Eun
DE
D052157:Aptamers, Nucleotide / Q000008:administration & dosage / Q000235:genetics*; D020868:Gene Silencing; D043583:Nucleoside-Triphosphatase / Q000037:antagonists & inhibitors / Q000235:genetics*; D020365:RNA Helicases / Q000235:genetics*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000502:physiology*
D052157:Aptamers, Nucleotide; D043583:Nucleoside-Triphosphatase; D020365:RNA Helicases