Rational optimization of the DSL ligase ribozyme with GNRA/receptor interacting modules.
The DSL ribozyme is a class of artificial ligase ribozymes with a highly modular architecture, which catalyzes template-directed RNA ligation on a helical substrate module that can be either covalently connected (cis-DSL) or physically separated (trans-DSL) from the catalytic module. Substrate recognition by the catalytic module is promoted by one or two sets of GNRA/receptor interactions acting as clamps in the cis or trans configurations, respectively. In this study, we have rationally designed and analyzed the catalytic and self-assembly properties of several trans-DSL ribozymes with different sets of natural and artificial GNRA-receptor clamps. Two variants newly designed in this study showed significantly enhanced catalytic properties with respect of the original trans-DSL construct. While this work allows dissection of the turnover and catalytic properties of the trans-DSL ribozyme, it also emphasizes the remarkable modularity of RNA tertiary structure for nano-construction of complex functions.
Ishikawa
Junya
J
Department of Chemistry and Biochemistry, Graduate School of Engineering, Kyushu University, Fukuoka 819-0395, Japan.
Matsumura
Shigeyoshi
S
Jaeger
Luc
L
Inoue
Tan
T
Furuta
Hiroyuki
H
Ikawa
Yoshiya
Y
D001483:Base Sequence; D001698:Biomedical Engineering; D020134:Catalytic Domain; D015195:Drug Design; D007700:Kinetics; D009690:Nucleic Acid Conformation; D012313:RNA / Q000235:genetics / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013237:Stereoisomerism; D013379:Substrate Specificity
D016337:RNA, Catalytic; D012313:RNA