Substitution of non-catalytic stem and loop regions of hammerhead ribozyme with DNA counterparts only increases KM without sacrificing the catalytic step (kcat): a way to improve substrate-specificity.
In elucidating structure-function relationships and stabilizing ribozymes in vivo, several chimeric RNA/DNA ribozymes and substrates were chemically synthesized. Measurements of kinetic parameters revealed that the maximally deoxyribonucleotide-substituted ribozyme (DRDRD32) gained the highest catalytic activity reaching the kcat value of > 10 min-1, the highest value ever reported for hammerhead-type ribozymes. Since these chimeric ribozymes are more stable than the wild-type all-RNA ribozymes in vivo and they also possess higher substrate-specificity, they are considered to be better candidates for antiviral therapeutic agents.
Shimayama
T
T
Hitachi Chemical Co., Tsukuba Science City, Japan.
Sawata
S
S
Komiyama
M
M
Takagi
Y
Y
Tanaka
Y
Y
Wada
A
A
Sugimoto
N
N
Rossi
J J
JJ
Nishikawa
F
F
Nishikawa
S
S
D001483:Base Sequence; D002384:Catalysis; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism*; D012260:Ribonucleases / Q000378:metabolism; D013379:Substrate Specificity
D016337:RNA, Catalytic; D012313:RNA; D004247:DNA; D012260:Ribonucleases