Conserved base-pairings between C266-A268 and U307-G309 in the P7 of the Tetrahymena ribozyme is nonessential for the in vitro self-splicing reaction.
P7 is highly conserved in Group I self-splicing intron ribozymes. This region is known to coordinate metal ions and bind a cofactor guanosine required for the self-splicing. To further investigate the fundamental role of the corresponding region in the Tetrahymena ribozyme, we attempted to identify minimal requirements for the base-paired region excluding the guanosine binding site. We discovered that a variety of sequences are eligible and its derivatives possessing extra nucleotide(s) can still conduct the first step of splicing, indicating that no particular base-pairing is essential in this region for conducting the reaction in vitro. The results provide two hypotheses for the fundamental role of this region: (i) if the region contains element(s) that are strictly required in the catalysis, they are not necessarily tightly fixed in the ribozyme and (ii) if not, its fundamental role may simply be to coordinate neighboring regions that are directly involved in the catalysis.
Oe
Y
Y
Graduate School of Science, Kyoto University, Kyoto, 606-8502, Japan.
Ikawa
Y
Y
Shiraishi
H
H
Inoue
T
T
D000818:Animals; D020029:Base Pairing; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D005091:Exons; D007438:Introns; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000737:chemistry* / Q000378:metabolism*; D013768:Tetrahymena / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D016337:RNA, Catalytic; D008274:Magnesium