Lazowska J, Meunier B, Macadre C
Centre de Génétique Moléculaire du CNRS, Gif-sur-Yvette, France.
EMBO J. 1994 Oct 17;13(20):4963-72. doi: 10.1002/j.1460-2075.1994.tb06823.x.
Group II introns ai1 and ai2 of the Saccharomyces cerevisiae mitochondrial COXI gene encode proteins having a dual function (maturase and reverse transcriptase) and are mobile genetic elements. By construction of adequate donor genomes, we demonstrate that each of them is self-sufficient and practises homing in the absence of homing-type endonucleases encoded by either group I introns or the ENS2 gene. Each of the S. cerevisiae group II self-mobile introns was tested for its ability to invade mitochondrial DNA (mtDNA) from two related Saccharomyces species. Surprisingly, only ai2 was observed to integrate into both genomes. The non-mobility of ai1 was clearly correlated with some polymorphic changes occurring in sequences flanking its insertion sites in the recipient mtDNAs. Importantly, studies of the behaviour of these introns in interspecific crosses demonstrate that flanking marker co-conversion accompanying group II intron homing is unidirectional and efficient only in the 3' to 5' direction towards the upstream exon. Thus, the polar co-conversion and dependence of the splicing proficiency of the intron reported previously by us are hallmarks of group II intron homing, which significantly distinguish it from the strictly DNA-based group I intron homing and strictly RNA-based group II intron transposition.
酿酒酵母线粒体COXI基因的II组内含子ai1和ai2编码具有双重功能(成熟酶和逆转录酶)的蛋白质,并且是可移动遗传元件。通过构建合适的供体基因组,我们证明它们各自是自给自足的,并且在没有由I组内含子或ENS2基因编码的归巢型内切核酸酶的情况下进行归巢。对酿酒酵母的每个II组自移动内含子进行了测试,以检测其从两个相关酿酒酵母物种侵入线粒体DNA(mtDNA)的能力。令人惊讶的是,仅观察到ai2整合到两个基因组中。ai1的不可移动性显然与受体mtDNA中其插入位点侧翼序列中发生的一些多态性变化相关。重要的是,对这些内含子在种间杂交中行为的研究表明,伴随II组内含子归巢的侧翼标记共转化是单向的,并且仅在朝着上游外显子的3'至5'方向上有效。因此,我们之前报道的内含子剪接熟练程度的极性共转化和依赖性是II组内含子归巢的标志,这使其与基于严格DNA的I组内含子归巢和基于严格RNA的II组内含子转位显著区分开来。