Moran J V, Wernette C M, Mecklenburg K L, Butow R A, Perlman P S
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235-9038.
Nucleic Acids Res. 1992 Aug 11;20(15):4069-76. doi: 10.1093/nar/20.15.4069.
We have found that intron 5 alpha of the COXI gene (al5 alpha) of yeast mtDNA is a mobile group I intron in crosses between strains having or lacking the intron. We have demonstrated the following hallmarks of that process: 1) co-conversion of flanking optional intron markers; 2) mutations that truncate the intron open reading frame block intron mobility; and 3) the intron open reading frame encodes an endonuclease activity that is required for intron movement. The endonuclease activity, termed I-Sce IV, cleaves the COXI allele lacking al5 alpha near the site of intron insertion, making a four-base staggered cut with 3' OH overhangs. Three cloned DNAs derived from different forms of the COXI gene, which differ in primary sequence at up to seven nucleotides around the cleavage site, are all good substrates for in vitro I-Sce IV cleavage activity. Two of the strains from which these substrates were derived were tested in crosses and are comparably efficient as al5 alpha recipients. When compared with omega mobility occurring simultaneously in one cross, al5 alpha is less efficient as a mobile element.
我们发现酵母线粒体DNA的COXI基因的内含子5α(al5α)在有或没有该内含子的菌株杂交中是一种可移动的I类内含子。我们已经证明了该过程的以下特征:1)侧翼可选内含子标记的共转化;2)截断内含子开放阅读框的突变会阻止内含子移动;3)内含子开放阅读框编码内含子移动所需的内切核酸酶活性。这种内切核酸酶活性称为I-Sce IV,在缺乏al5α的COXI等位基因的内含子插入位点附近切割,产生具有3' OH突出端的四碱基交错切口。来自COXI基因不同形式的三个克隆DNA,在切割位点周围多达七个核苷酸的一级序列上有所不同,都是体外I-Sce IV切割活性的良好底物。从中获得这些底物的两个菌株在杂交中进行了测试,并且作为al5α受体具有相当的效率。与在一次杂交中同时发生的ω移动性相比,al5α作为可移动元件的效率较低。