Lambert J R, Bilanchone V W, Cumsky M G
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7345-9. doi: 10.1073/pnas.91.15.7345.
The yeast COX5a and COX5b genes encode isoforms of subunit Va of the mitochondrial inner membrane protein complex cytochrome c oxidase. These genes have been shown to be inversely regulated at the level of transcription by oxygen, which functions through the metabolic coeffector heme. In earlier studies we identified several regulatory elements that control transcriptional activation and aerobic repression of one of these genes, COX5b. Here, we report the isolation of trans-acting mutants that are defective in the aerobic repression of COX5b transcription. The mutants fall into two complementation groups. One group specifies ROX1, which encodes a product reported to be involved in transcriptional repression. The other group identified the gene we have designated ORD1. Mutations in ORD1 cause overexpression of COX5b aerobically but do not affect the expression of the hypoxic genes CYC7, HEM13, and ANB1. ORD1 mutations also do not affect the expression of the aerobic genes COX5a, CYC1, ROX1, ROX3, and TIF51A. The yeast genome contains a single ORD1 gene that resides on chromosome XI. Strains carrying chromosomal deletions of the ORD1 locus are viable and exhibit phenotypes similar to, but less severe than, that of the original mutant. The nucleotide sequence of ORD1 revealed that it is identical to IXR1, a yeast gene whose product contains two high mobility group boxes, binds to platinated DNA, and confers sensitivity to the antitumor drug cisplatin. Consistent with the latter observations, we found that the ORD1 product could bind to both the upstream region of COX5b and to DNA modified with cisplatin.
酵母COX5a和COX5b基因编码线粒体内膜蛋白复合物细胞色素c氧化酶亚基Va的同工型。这些基因已被证明在转录水平上受氧气的反向调节,氧气通过代谢效应物血红素发挥作用。在早期研究中,我们鉴定了几个控制其中一个基因COX5b转录激活和有氧抑制的调控元件。在此,我们报告了在COX5b转录的有氧抑制中存在缺陷的反式作用突变体的分离。这些突变体分为两个互补组。一组确定为ROX1,其编码一种据报道参与转录抑制的产物。另一组鉴定出我们命名为ORD1的基因。ORD1中的突变导致COX5b在有氧条件下过度表达,但不影响缺氧基因CYC7、HEM13和ANB1的表达。ORD1突变也不影响有氧基因COX5a、CYC1、ROX1、ROX3和TIF51A的表达。酵母基因组包含一个位于第十一条染色体上的单一ORD1基因。携带ORD1基因座染色体缺失的菌株是有活力的,并且表现出与原始突变体相似但程度较轻的表型。ORD1的核苷酸序列显示它与IXR1相同,IXR1是一个酵母基因,其产物包含两个高迁移率族框,与铂化DNA结合,并赋予对抗肿瘤药物顺铂的敏感性。与后一种观察结果一致,我们发现ORD1产物可以与COX5b的上游区域以及用顺铂修饰的DNA结合。