Self-splicing of group II introns in vitro: Lariat formation and 3′ splice site selection in mutant RNAs

Abstract

Deletion or substitution of the branch A residue in group II intron bl1 significantly reduces splicing activity; yet, residual exon ligation is correct, and lariats have their branch points at the normal distance from the 3′ end of the intron. Mutations in the sequence facing the branch point also allow residual lariat formation; however, free 3′ exons are generated with false 5′ termini, all of which are within a UCACA consensus sequence located upstream or downstream of the normal 3′ splice site. These results indicate that both the conserved 3′ splice site APy and the spatial arangements in stem 6 are crucial for correct 3′ splice site selection.