Modifying in vitro reaction conditions for group II intron self-splicing reveals novel reactions including spliced exon reopening.
Reveals novel RNA reactions in vitro; leaves open physiological relevance and requires cellular validation before mechanistic extension.
Reaction parameters were modified to enhance the in vitro reaction rate and to reveal partial and novel reactions of the group II intron 5g of the mitochondrial gene from Saccharomyces cerevisiae encoding cytochrome c oxidase subunit I. One alteration yields separate 5'- and 3'-exons plus linear excised intron as the main products. A linear reaction intermediate, containing intron and 3'-exon, and products resulting from cleavages at two unexpected sites were identified. Spliced exon "reopening," a novel reaction between excised intron and spliced exons, appears responsible for separate 5'- and 3'-exon products.
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Jarrell et al. (1988) studied this question.
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