Intermolecular exon ligation of the rRNA precursor of Tetrahymena: oligonucleotides can function as 5' exons

Abstract

The dinucleotide CpUOH, when incubated with self-splicing Tetrahymena pre-rRNA in the absence of GTP, functions as a 5' exon. It cleaves the precursor exactly at the 3' splice site and becomes covalently ligated to the 3' exon. Other oligonucleotides with sequences that resemble CUCUCU, the sequence at the 3' end of the 5' exon, can add to the 3' exon in this reaction. Such splicing in trans is most readily explained by a site within the intervening sequence that binds the last few nucleotides of the 5' exon. This binding site functions in splice site recognition and is also part of the active site of the ribozyme. The mechanism by which 5' splice sites are selected in Tetrahymena rRNA and group I mitochondrial RNA splicing is like that used in nuclear mRNA splicing, in that it involves specific pairing of bases adjacent to the splice site with a complementary RNA sequence.