Properties of antigenomic hepatitis delta virus ribozyme that consists of three RNA oligomer strands.
A three-strand ribozyme, a derivative of antigenomic hepatitis delta virus (HDV) ribozyme, which consists of subfragments of 16 (L), 17 (S), and 33 nucleotides (B), has been constructed. The ternary B-L-S complex formed by the subfragments in stoichiometric ratio was able to catalyze a self-cleavage reaction. Kinetics of this reaction exhibited biphasic behavior and the same parameters as in the case of natural cis-ribozyme. Study of kinetics of reaction initiated by adding various reaction components and the study of binary complex formation between subfragments B and L, B and S, and also ternary B-L-S complex formation revealed that: 1) in the presence of Mg2+, B and S form a stoichiometric complex, L and S do not form complex at all, while B and L form 2 types of complexes, probably B-L and 2B-L; and addition of S subfragment prevented the formation of the latter complex; 2) the reaction initiated by S subfragment proceeds much slower than that initiated by other components pointing to the possibility that in the absence of S L may form a nonproductive complex with B, which is slowly displaced by S followed by productive ternary complex formation. Dissociation constants for binary B-L, B-S and ternary B-L-S complexes have been estimated.
Belianko
T I
TI
Russian Cardiology Research and Development Complex, Russian Academy of Medical Sciences, Moscow, 121552, Russia. lasav@cardio.ru
Alekseenkova
V A
VA
Savochkina
L P
LP
Lukin
M A
MA
Beabealashvilli
R Sh
RSh
D001483:Base Sequence / Q000235:genetics; D002384:Catalysis; D016679:Genome, Viral; D003698:Hepatitis Delta Virus / Q000235:genetics*; D008274:Magnesium / Q000737:chemistry; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics*; D016337:RNA, Catalytic / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D012367:RNA, Viral; D013329:Structure-Activity Relationship
D016372:RNA, Antisense; D016337:RNA, Catalytic; D012367:RNA, Viral; D008274:Magnesium