Marykwas D L, Fox T D
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Mol Cell Biol. 1989 Feb;9(2):484-91. doi: 10.1128/mcb.9.2.484-491.1989.
The product of the Saccharomyces cerevisiae nuclear gene PET494 is required to promote the translation of the mitochondrial mRNA encoding cytochrome c oxidase subunit III (coxIII). The level of cytochrome c oxidase activity is affected by several different environmental conditions, which also influence coxIII expression. We have studied the regulation of PET494 to test whether the level of its expression might modulate coxIII translation in response to these conditions. A pet494::lacZ fusion was constructed and used to monitor PET494 expression. PET494 was regulated by oxygen availability: expression in a respiration-competent diploid strain grown anaerobically was one-fifth the level of expression in aerobically grown cells. However, since PET494 mRNA levels did not vary in response to oxygen deprivation, regulation by oxygen appears to occur at the translational level. This oxygen regulation was not mediated by heme, and PET494 expression was independent of the heme activator protein HAP2. The regulation of PET494 expression by carbon source was also examined. In cells grown on glucose-containing medium, PET494 was expressed at levels four- to sixfold lower than in cells grown on ethanol and glycerol. However, addition of ethanol to glucose-containing medium induced PET494 expression approximately twofold. PET494 transcript levels varied over a fourfold range in response to different carbon sources. The effects on PET494 expression of mutations in the SNF1, SNF2, SSN6, and HXK2 genes were also determined and found to be minimal.
酿酒酵母核基因PET494的产物是促进编码细胞色素c氧化酶亚基III(coxIII)的线粒体mRNA翻译所必需的。细胞色素c氧化酶活性水平受几种不同环境条件的影响,这些条件也会影响coxIII的表达。我们研究了PET494的调控,以测试其表达水平是否可能根据这些条件调节coxIII的翻译。构建了pet494::lacZ融合体并用于监测PET494的表达。PET494受氧气供应的调控:在厌氧生长的具有呼吸能力的二倍体菌株中的表达是需氧生长细胞中表达水平的五分之一。然而,由于PET494 mRNA水平不会因缺氧而变化,氧气调控似乎发生在翻译水平。这种氧气调控不是由血红素介导的,并且PET494的表达独立于血红素激活蛋白HAP2。还研究了碳源对PET494表达的调控。在含葡萄糖培养基上生长的细胞中,PET494的表达水平比在乙醇和甘油上生长的细胞低四到六倍。然而,向含葡萄糖培养基中添加乙醇可使PET494的表达诱导约两倍。PET494转录水平因不同碳源而在四倍范围内变化。还确定了SNF1、SNF2、SSN6和HXK2基因突变对PET494表达的影响,发现影响很小。