[Reconstruction of hepatitis D virus ribozymes and their ability to trans-cleave hepatitis C virus RNA].
OBJECTIVE
To study whether reconstructed hepatitis D virus (HDV) ribozymes have the ability to trans-cleave hepatitis C virus (HCV) RNA.


METHODS
We reconstructed the stem IV and substrate-binding region of HDV genomic ribozymes, thus obtained three HDV ribozymes named RzC1, RzC2 and RzC3 expected to be able to trans-cleave HCV RNA at predicted sites. The substrate containing HCV RNA 5'-noncoding region (5'-NCR) and 5'-fragment of C region (HCV RNA 5'-NCR-C) was synthesized by transcription in vitro, then radiolabelled at its 5'-end. Under certain pH and appropriate concentration of Mg(2+) with or without deionized formamide, the ribozymes and their substrate were mixed at mol ratio of 100 : 1 and reacted for two hours. Trans-cleaved products were shown by denatured polyacrylamide gel electrophoresis and autoradiography, and the percentage of trans-cleaved substrate was calculated as the activity indicator of these ribozymes.


RESULTS
RzC1 and RzC2 were able to trans-cleave HCV RNA 5'-NCR-C site specifically, and their activity could be enhanced by certain concentration of deionized formamide. RzC3 could not trans-cleave the substrate.


CONCLUSIONS
Optimized HDV genomic ribozymes can trans-cleave HCV RNA.
Yu
Y
Y
Institute of Infectious Diseases of PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Mao
Q
Q
Li
Q
Q
Wang
Y
Y
Min
F
F
D020121:5' Untranslated Regions / Q000378:metabolism; D016174:Hepacivirus / Q000235:genetics*; D003698:Hepatitis Delta Virus / Q000201:enzymology*; D016337:RNA, Catalytic / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*
D020121:5' Untranslated Regions; D016337:RNA, Catalytic; D012367:RNA, Viral