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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >The Rieske FeS protein encoded and synthesized within mitochondria complements a deficiency in the nuclear gene.
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The Rieske FeS protein encoded and synthesized within mitochondria complements a deficiency in the nuclear gene.

机译：线粒体内编码和合成的Rieske FeS蛋白补充了核基因的缺陷。

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摘要

The Rieske FeS protein, an essential catalytic subunit of the mitochondrial cytochrome bc1 complex, is encoded in yeast by the nuclear gene RIP1, whose deletion leads to a respiratory-deficient phenotype. By using biolistic transformation, we have relocated the nuclear RIP1 gene into mitochondria. To allow its expression within the organelle and to direct its integration downstream of the cox1 gene, we have fused the 3' end of the Saccharomyces douglasii cox1 gene upstream of the mitochondrial copy of RIP1 (RIP1m) flanked by the Saccharomyces cerevisiae cox1 promoter and terminator regions. We show that RIP1m integrated between the cox1 and atp8 genes is mitotically stable and expressed, and it complements a deletion of the nuclear gene. Immunodetection experiments demonstrate that the mitochondrial genome containing RIP1m is able to produce the Rip1 protein in lower steady-state amounts than the wild type but still sufficient to maintain a functional cytochrome bc1 complex and respiratory competence to a RIP1-deleted strain. Thus, this recombined mitochondrial genome is a fully functional mitochondrial chromosome with an extended gene content. This successful mitochondrial expression of a nuclear gene essential for respiration can be viewed at the evolutionary level as an artificial reversal of evolutionary events.
机译：Rieske FeS蛋白是线粒体细胞色素bc1复合体的重要催化亚基，由核基因RIP1在酵母中编码，RIP1的缺失导致呼吸缺陷型。通过使用生物射弹转化，我们已经将核RIP1基因重新定位到了线粒体中。为了使其在细胞器中表达并指导其整合到cox1基因的下游，我们融合了Saccharomyces douglasii cox1基因的3'端，位于RIP1（RIP1m）线粒体拷贝的上游，两侧是酿酒酵母cox1启动子和终止子地区。我们显示，cox1和atp8基因之间整合的RIP1m是有丝分裂稳定的和表达的，并且它补充了核基因的缺失。免疫检测实验表明，包含RIP1m的线粒体基因组能够以比野生型更低的稳态量生产Rip1蛋白，但仍足以维持功能性细胞色素bc1复合物和对RIP1缺失菌株的呼吸能力。因此，这种重组的线粒体基因组是具有扩展基因含量的功能齐全的线粒体染色体。呼吸必不可少的核基因的这种线粒体成功表达可以在进化水平上视为进化事件的人为逆转。

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