Integration of group II intron bI1 into a foreign RNA by reversal of the self-splicing reaction in vitro.
Group II intron bI1, the first intron of the COB gene in the mitochondria of S. cerevisiae, is able to self-splice in vitro with the basic pathway similar to nuclear pre-mRNA splicing. We show that incubation of the intron lariat with ligated exons bE1 and bE2 leads to a complete reversal of the splicing reaction. The integration of the intron into the ligated exons is correct; the reconstituted preRNA of the reverse reaction can undergo a self-splicing reaction anew. When incubated with a foreign RNA species bearing a sequence motif that is complementary to exon binding site 1, the lariat can integrate into this RNA with the position of insertion immediately downstream of this sequence. This result implies that transposition of group II introns on the RNA level by reversal of the splicing reaction is, in principle, conceivable.
Institut für Genetik und Mikrobiologie, Universität München, Federal Republic of Germany.
M
M
Mörl
C
C
Schmelzer
D001483:Base Sequence; D004272:DNA, Mitochondrial / Q000235:genetics*; D005800:Genes, Fungal; D007438:Introns; D008969:Molecular Sequence Data; D012322:RNA Precursors / Q000235:genetics; D012326:RNA Splicing; D016337:RNA, Catalytic; D012335:RNA, Ribosomal / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014158:Transcription, Genetic
D004272:DNA, Mitochondrial; D012322:RNA Precursors; D016337:RNA, Catalytic; D012335:RNA, Ribosomal