Group II intron splicing in Escherichia coli:: phenotypes of cis-acting mutations resemble splicing defects observed in organelle RNA processing

The mitochondrial group IIB intron rl1, from the green algae Scenedesmus obliquus' LSUrRNB gene, has been introduced into the lacZ gene encoding beta-galactosidase, After DNA-mediated transformation of the recombinant lacZ gene into Escherichia coli, we observed correct splicing of the chimeric precursor RNA in vivo. In contrast to autocatalytic in vitro self-splicing, intron processing in vivo is independent of the growth temperature, suggesting that in E.coli, trans-acting factors are involved in group II intron splicing. Such a system would seem suitable as a model for analyzing intron processing in a prokaryotic host, In order to study further the effect of cis-mutations on intron splicing, different rl1 mutants were analyzed (with respect to their splicing activity) in E.coli. Although the phenotypes of these E.coli intron splicing mutants were identical to those which can be observed during organellar splicing of rl1, they are different to those observed in in vitro self-splicing experiments. Therefore, in both organelles and prokaryotes, it is likely that either similar splicing factors or trans-acting factors exhibiting similar functions are involved in splicing. We speculate that ubiquitous trans-acting factors, via recent horizontal transfer, have contributed to the spread of group II introns.