McEwen J E, Hong K H, Park S, Preciado G T
Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90024-1489.
Curr Genet. 1993 Jan;23(1):9-14. doi: 10.1007/BF00336742.
The nuclear genes PET117 and PET191 are required for the assembly of active cytochrome c oxidase in S. cerevisiae, yet their gene products are not subunits of the final assembled cytochrome c oxidase complex. Plasmids bearing PET117 or PET191 were isolated by their ability to complement the pet117-1 or pet191-1 mutations, respectively. By restriction mapping, subcloning, and deletion analysis of yeast DNA fragments that complement these mutations, the PET117 and PET191 genes were localized to smaller regions of DNA, which were then sequenced from both strands. The PET117 open reading frame is of 107 codons and the PET191 open reading frame is of 108 codons. Neither the PET191 nor PET117 DNA sequences have been reported previously, and the derived amino-acid sequences of the PET191 and PET117 open reading frames exhibit no significant primary amino-acid sequence similarity to other protein sequences available in the NBRF data base, or from translated Genbank sequences. By hybridization of PET117 or PET191 probes first to a chromosome blot and next to a library of physically mapped fragments of yeast genomic DNA, the map locations of the PET191 and PET117 genes were determined. PET117 is located on chromosome V near the HIS1 gene and PET191 is located on chromosome X near the CYC1 gene.
核基因PET117和PET191是酿酒酵母中活性细胞色素c氧化酶组装所必需的,但其基因产物并非最终组装的细胞色素c氧化酶复合物的亚基。分别通过其互补pet117 - 1或pet191 - 1突变的能力,分离出携带PET117或PET191的质粒。通过对互补这些突变的酵母DNA片段进行限制性图谱分析、亚克隆和缺失分析,将PET117和PET191基因定位到更小的DNA区域,然后从两条链对其进行测序。PET117开放阅读框有107个密码子,PET191开放阅读框有108个密码子。PET191和PET117的DNA序列以前均未被报道,PET191和PET117开放阅读框推导的氨基酸序列与NBRF数据库中或翻译后的Genbank序列中的其他蛋白质序列没有显著的一级氨基酸序列相似性。通过首先将PET117或PET191探针与染色体印迹杂交,然后与酵母基因组DNA物理图谱片段文库杂交,确定了PET191和PET117基因的图谱位置。PET117位于第五条染色体上靠近HIS1基因的位置,PET191位于第十条染色体上靠近CYC1基因的位置。