Skelly P J, Clark-Walker G D
Research School of Biological Sciences, Australian National University, Canberra.
Mol Cell Biol. 1990 Apr;10(4):1530-7. doi: 10.1128/mcb.10.4.1530-1537.1990.
Saccharomyces cerevisiae mitochondrial DNA deletion mutants have been used to examine whether base-biased intergenic regions of the genome influence mitochondrial biogenesis. One strain (delta 5.0) lacks a 5-kilobase (kb) segment extending from the proline tRNA gene to the small rRNA gene that includes ori1, while a second strain (delta 3.7) is missing a 3.7-kb region between the genes for ATPase subunit 6 and glutamic acid tRNA that encompasses ori7 plus ori2. Growth of these strains on both fermentable and nonfermentable substrates does not differ from growth of the wild-type strain, indicating that the deletable regions of the genome do not play a direct role in the expression of mitochondrial genes. Examination of whether the 5- or 3.7-kb regions influence mitochondrial DNA transmission was undertaken by crossing strains and examining mitochondrial genotypes in zygotic colonies. In a cross between strain delta 5.0, harboring three active ori elements (ori2, ori3, and ori5), and strain delta 3.7, containing only two active ori elements (ori3 and ori5), there is a preferential recovery of the genome containing two active ori elements (37% of progeny) over that containing three active elements (20%). This unexpected result, suggesting that active ori elements do not influence transmission of respiratory-competent genomes, is interpreted to reflect a preferential conversion of the delta 5.0 genome to the wild type (41% of progeny). Supporting evidence for conversion over biased transmission is shown by preferential recovery of a nonparental genome in the progeny of a heterozygous cross in which both parental molecules can be identified by size polymorphisms.
酿酒酵母线粒体DNA缺失突变体已被用于研究基因组中碱基偏向的基因间区域是否影响线粒体生物发生。一个菌株(delta 5.0)缺失了一个从脯氨酸tRNA基因延伸到小rRNA基因的5千碱基(kb)片段,该片段包括ori1,而第二个菌株(delta 3.7)在ATPase亚基6基因和谷氨酸tRNA基因之间缺失了一个3.7 kb的区域,该区域包含ori7加ori2。这些菌株在可发酵和不可发酵底物上的生长与野生型菌株的生长没有差异,这表明基因组的可缺失区域在线粒体基因的表达中不发挥直接作用。通过杂交菌株并检查合子菌落中的线粒体基因型,研究了5 kb或3.7 kb区域是否影响线粒体DNA传递。在含有三个活性ori元件(ori2、ori3和ori5)的delta 5.0菌株与仅含有两个活性ori元件(ori3和ori5)的delta 3.7菌株之间的杂交中,与含有三个活性元件的基因组(20%的后代)相比,含有两个活性ori元件的基因组(37%的后代)有优先恢复。这个意外的结果表明活性ori元件不影响具有呼吸能力的基因组的传递,被解释为反映了delta 5.0基因组向野生型的优先转化(41%的后代)。在杂合杂交的后代中优先恢复一个非亲本基因组,显示了支持转化而非偏向传递的证据,在该杂交中,两个亲本分子可以通过大小多态性来识别。