Hepatitis C virus (HCV), a hepacivirus member of the Flaviviridae family, has a positive-stranded RNA genome, which consists of a single open reading frame (ORF) and nontranslated regions (NTRs) at the 5′ and 3′ ends. The 5′NTR was found to contain an internal ribosomal entry site (IRES), which is required for the translation of HCV mRNA. Moreover, the 5′NTR is likely to play a key role in the replication of viral RNA. To identify the cis-acting element required for viral RNA replication, chimeric subgenomic replicons of HCV were generated. Dissection of the replication element from the translation element was accomplished by inserting the polioviral IRES between the serially deleted 5′NTR of HCV and ORF encoding neomycin phosphotransferase. The deletions of the 5′NTR of HCV were performed according to the secondary structure of HCV. Replicons containing domains I and II supported RNA replication and further deletion toward the 5′ end abolished replication. The addition of domain III and the pseudoknot structure of the 5′NTR to domains I and II augmented the colony-forming efficiency of replicons by 100-fold. This indicates that domains I and II are necessary and sufficient for replication of RNA and that almost all of the 5′NTR is required for efficient RNA replication.
|Number of pages||8|
|Journal||Biochemical and biophysical research communications|
|Publication status||Published - 2002|
- Cis-acting element
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology