Mobility of yeast mitochondrial group II introns: engineering a new site specificity and retrohoming via full reverse splicing

Article Abstract:

Enzyme assays and nucleic acid manipulation techniques were used to investigate the mobility of yeast mitochondrial group II introns, al1 and al2. The results showed that sense-strand cleavage for al2 occurs mainly through a partial reverse splicing reaction while that for al1 entails complete reverse splicing which results to the insertion of the linear intron RNA into double-stranded DNA. It was also found that al1 homing and reverse splicing is dependent on the EBS1(RNA)/IBS1(DNA) pairing and that target specificity can be altered by compensatory changes in the target site and the donor intron. Homing was documented to proceed via RT-dependent and -independent pathways.

Methods, Analysis, Usage, RNA, Molecular biology, Yeast, Yeast (Food product), RNA splicing, Molecular genetics, Enzymatic analysis

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Reverse transcriptase activity associated with maturase-encoding group II introns in yeast mitochondria

Article Abstract:

Effective techniques for studying reverse transcription in Neurospora mitochondria are used to show that yeast mitochrondria have a reverse transcriptase (RT) activity that is likely encoded by and highly specific for the group II introns al1 and al2, and their flanking exons. Thus, these introns are possibly retroelements, which encode RTs that specifically copy the intron. In effect, the retroelements are able to adapt to function in splicing through their ability to bind to the intron in unspliced precursor RNA.

Mitochondria

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Group II intron mobility occurs by target DNA-primed reverse transcription

Article Abstract:

The homing of the yeast mtDNA group II intron al2 takes place by reverse transcriptase at a double-strand break in the recipient DNA. Reverse transcription of al2-containing pre-mRNA is primed by the antisense strand cleaved in exon 3 and causes cotransfer of the intron and flanking exon sequences. Interestingly, the DNA endonuclease which initiates homing requires both the al2 reverse transcriptase protein and al2RNA.

Mitochondrial DNA, Reverse transcriptase, Exon (Molecular genetics), Exons (Molecular genetics)

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