Generation of a catalytic module on a self-folding RNA.
It is theoretically possible to obtain a catalytic site of an artificial ribozyme from a random sequence consisting of a limited numbers of nucleotides. However, this strategy has been inadequately explored. Here, we report an in vitro selection technique that exploits modular construction of a structurally constrained RNA to acquire a catalytic site for RNA ligation from a short random sequence. To practice the selection, a sequence of 30 nucleotides was located close to the putative reaction site in a derivative of a naturally occurring self-folding RNA whose crystal structure is known. RNAs whose activity depended on the starting three-dimensional structure were selected with 3'-5' ligation specificity, indicating that the strategy can be used to acquire a variety of catalytic sites and other functional RNA modules.
Yoshioka
Wataru
W
Graduate School of Biostudies, Kyoto University, 606-8502 Japan.
Ikawa
Yoshiya
Y
Jaeger
Luc
L
Shiraishi
Hideaki
H
Inoue
Tan
T
D000818:Animals; D001483:Base Sequence; D020134:Catalytic Domain; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013768:Tetrahymena / Q000235:genetics / Q000378:metabolism
D016337:RNA, Catalytic; D016053:RNA, Protozoan; D012313:RNA