Coffin J W, Dhillon R, Ritzel R G, Nargang F E
Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
Curr Genet. 1997 Oct;32(4):273-80. doi: 10.1007/s002940050277.
The cya-5 nuclear mutant of Neurosopora crassa was previously shown to be deficient in cytochrome aa3, cytochrome c oxidase activity, and the immunologically detectable COXI protein. We have now demonstrated that the mitochondria of this mutant contain mRNA for the COXI protein and that COXI cannot be detected during pulse-chase labeling experiments of mitochondrial translation products. Cloning and analysis of the cya-5 gene reveal a long open reading frame capable of encoding a 1136 amino-acid protein. Sequence analysis suggests that the potential CYA-5 protein contains a mitochondrial targeting sequence at its amino-terminus. The long open reading frame also contains a 200 amino-acid region with homology to the PET309 protein, which is required for the production or stability of intron-containing coxI mRNAs, as well as the translation of mature coxI mRNAs, in the yeast Saccharomyces cerevisiae. These data suggest that the CYA-5 protein of N. crassa is required in a post-transcriptional step for COXI expression, most probably for the efficient translation of coxI mRNA.
粗糙脉孢菌的cya - 5核突变体先前已被证明缺乏细胞色素aa3、细胞色素c氧化酶活性以及免疫可检测的COXI蛋白。我们现在已经证明,该突变体的线粒体含有COXI蛋白的mRNA,并且在脉冲追踪标记线粒体翻译产物的实验中无法检测到COXI。对cya - 5基因的克隆和分析揭示了一个能够编码1136个氨基酸的长开放阅读框。序列分析表明,潜在的CYA - 5蛋白在其氨基末端含有一个线粒体靶向序列。这个长开放阅读框还包含一个与PET309蛋白具有同源性的200个氨基酸区域,PET309蛋白对于酿酒酵母中含内含子的coxI mRNA的产生或稳定性以及成熟coxI mRNA的翻译是必需的。这些数据表明,粗糙脉孢菌的CYA - 5蛋白在COXI表达的转录后步骤中是必需的,很可能是coxI mRNA有效翻译所必需的。