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PMID:9118229
| Citation |
Eskes, R, Yang, J, Lambowitz, AM and Perlman, PS (1997) Mobility of yeast mitochondrial group II introns: engineering a new site specificity and retrohoming via full reverse splicing. Cell 88:865-74 |
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| Abstract |
The mobile group II introns aI1 and aI2 of yeast mtDNA encode endonuclease activities that cleave intronless DNA target sites to initiate mobility by target DNA-primed reverse transcription. For aI2, sense-strand cleavage occurs mainly by a partial reverse splicing reaction, whereas for aI1, complete reverse splicing occurs, leading to insertion of the linear intron RNA into double-stranded DNA. Here, we show that aI1 homing and reverse splicing depend on the EBS1 (RNA)/IBS1(DNA) pairing and that target specificity can be changed by compensatory changes in the target site and the donor intron. Using well-marked strains to follow coconversion of flanking DNA, we show that homing occurs by both RT-dependent and -independent pathways. Remarkably, in most RT-dependent events, the reverse spliced intron is the initial template for first-strand cDNA synthesis. |
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| Keywords |
Base Sequence; DNA, Complementary/analysis; DNA, Complementary/chemical synthesis; DNA, Fungal/analysis; DNA, Mitochondrial/physiology; Endonucleases/genetics; Gene Expression Regulation, Enzymologic/genetics; Gene Expression Regulation, Fungal/genetics; Introns/genetics; Mutagenesis, Site-Directed/genetics; Plasmids; RNA Splicing/genetics; Recombination, Genetic; Yeasts/enzymology; Yeasts/genetics |
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Significance
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