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Mobile group II introns of yeast mitochondrial DNA are novel site-specific retroelements.

机译:酵母线粒体DNA的流动II类内含子是新型的位点特异性逆转录元件。

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摘要

Group II introns aI1 and aI2 of the yeast mitochondrial COXI gene are mobile elements that encode an intron-specific reverse transcriptase (RT) activity. We show here that the introns of Saccharomyces cerevisiae ID41-6/161 insert site specifically into intronless alleles. The mobility is accompanied by efficient, but highly asymmetric, coconversion of nearby flanking exon sequences. Analysis of mutants shows that the aI2 protein is required for the mobility of both aI1 and aI2. Efficient mobility is dependent on both the RT activity of the aI2-encoded protein and a separate function, a putative DNA endonuclease, that is associated with the Zn2+ finger-like region of the intron reading frame. Surprisingly, there appear to be two mobility modes: the major one involves cDNAs reverse transcribed from unspliced precursor RNA; the minor one, observed in two mutants lacking detectable RT activity, appears to involve DNA level recombination. A cis-dominant splicing-defective mutant of aI2 continues to synthesize cDNAs containing the introns but is completely defective in both mobility modes, indicating that the splicing or the structure of the intron is required. Our results demonstrate that the yeast group II intron aI2 is a retroelement that uses novel mobility mechanisms.
机译:酵母线粒体COXI基因的第II组内含子aI1和aI2是编码内含子特异性逆转录酶(RT)活性的移动元件。我们在这里显示,酿酒酵母ID41-6 / 161的内含子将位点插入到无内含子等位基因中。迁移性伴随着附近侧翼外显子序列的高效但高度不对称的共转化。突变体分析表明,aI2蛋白是aI1和aI2迁移所必需的。有效的迁移率既依赖于aI2编码蛋白的RT活性,又依赖于与内含子阅读框的Zn2 +手指状区域相关的独立功能,即推定的DNA内切核酸酶。出人意料的是,似乎有两种迁移模式:主要的一种涉及从未剪接的前体RNA逆转录的cDNA。在两个缺乏可检测的RT活性的突变体中观察到的次要突变体似乎与DNA水平重组有关。 aI 2的顺式为剪接缺陷的突变体继续合成含有内含子的cDNA,但是在两种迁移模式下都是完全缺陷的,表明需要内含子的剪接或结构。我们的结果表明,酵母II组内含子aI2是一种使用新型迁移机制的逆向元件。

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