We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
RNA Aptamers Targeted to Domain II of Hepatitis C Virus IRES That Bind to Its Apical Loop Region.
- Authors
Kikuchi, Kunio; Umehara, Takuya; Fukuda, Kotaro; Joonsung Hwang; Kuno, Atsushi; Hasegawa, Tsunemi; Nishikawa, Satoshi
- Abstract
The internal ribosome entry site (IRES) is important for translation of hepatitis C virus (HCV) mRNA and has a unique RNA structure containing conserved domains I to IV. To investigate the function of domain II, we selected RNA aptamers that bind to domain II of HCV IRES by applying a simple and convenient selection method using a hybridized tag for fixing domain II RNA on magnetic beads instead of synthesizing long RNA. In addition, we employed surface plasmon resonance (SPR) technology to measure the binding affinity of each generation and to obtain detailed kinetic constants. The selected aptamers have a consensus sequence, 5′-UAUGGCU-3′, which is complementary to the apical loop of domain II. The loop–loop interaction between the consensus sequence and domain II was confirmed by mutagenesis and nuclease mapping analyses. Binding affinities were dependent on the local structure containing the conserved sequence. The aptamers could inhibit IRES-dependent translation.
- Subjects
HEPATITIS C virus; RIBOSOMES; RNA; SURFACE plasmon resonance; MUTAGENESIS
- Publication
Journal of Biochemistry, 2003, Vol 133, Issue 3, p263
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/jb/mvg036