Properties of antigenomic hepatitis delta virus ribozyme cis- and trans- analogs.
A series of permuted variants of antigenomic HDV ribozyme and trans-acting variants were constructed. The catalytic activity study of the ribozymes has shown that all the variants were capable of self-cleaving with equally biphasic kinetics. Ribonuclease and Fe(II)-EDTA cleavage have provided evidence that all designed ribozymes fold according to the pseudoknot model and the conformations of the initial and cleaved ribozyme are different. A scheme of HDV ribozyme self-cleavage reaction was suggested. The role of hydrogen bonds in the reaction was evaluated by substitution of ribose in the ribozyme for deoxyribose. It was found that the 2'-OH group of U23 and C27 is critical for the reaction to occur; the 2'-OH group of U32 and U39 is important, while 2'-OH groups of other nucleotides of loop 3, stem 4 and stem 1 are unimportant for the cleavage activity.
Savochkina
L
L
Russian Cardiology Research and Development Center, Moscow, Russia. lasav@cardio.ru
Alekseenkova
V
V
Belyanko
T
T
Lukin
M
M
Beabealashvilli
R
R
D001483:Base Sequence; D002384:Catalysis; D016679:Genome, Viral; D003698:Hepatitis Delta Virus / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D016337:RNA, Catalytic / Q000737:chemistry*
D016337:RNA, Catalytic