INTERACTION OF INTRONIC BOUNDARIES IS REQUIRED FOR THE 2ND SPLICING STEP EFFICIENCY OF A GROUP-II INTRON

Group II and nuclear pre-mRNAs introns share a common splicing pathway involving a lariat intermediate, as well as some primary sequence similarities at the splice junctions. In this work, we analyze the role of the conserved nucleotides at the first and penultimate positions (G1 and A886) of a group II self-splicing intron. We show that the G1 nucleotide is essential for the efficiency of both the first and the second splicing steps, while substitutions at the penultimate nucleotide affect mostly the efficiency of the second step. A reciprocal suppression of the second splicing step defect can be observed in some double mutants. This result is best explained by a non-Watson - Crick interaction between the first and the penultimate nucleotides of the intron, which occurs after lariat formation. The finding that an interaction between intron boundaries is required for the second splicing step in both group II and nuclear pre-mRNA introns strengthens the idea that both systems employ similar mechanisms, albeit with differences in the details of the nucleotide interactions.