Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de Alimentos (IATA), Consejo Superior de Investigaciones Científicas (CSIC), Agustín Escardino 7, Paterna 46980, Valencia, Spain.
Departamento de Bioquímica y Biología Molecular, Universitat de València, Doctor Moliner 50, Burjassot 46100, Valencia, Spain; ERI Biotecmed, Universitat de València, Doctor Moliner 50, Burjassot 46100, Valencia, Spain.
Biochim Biophys Acta Gene Regul Mech. 2020 Jul;1863(7):194522. doi: 10.1016/j.bbagrm.2020.194522. Epub 2020 Mar 6.
Eukaryotic ribonucleotide reductases are iron-dependent enzymes that catalyze the rate-limiting step in the de novo synthesis of deoxyribonucleotides. Multiple mechanisms regulate the activity of ribonucleotide reductases in response to genotoxic stresses and iron deficiency. Upon iron starvation, the Saccharomyces cerevisiae Aft1 transcription factor specifically binds to iron-responsive cis elements within the promoter of a group of genes, known as the iron regulon, activating their transcription. Members of the iron regulon participate in iron acquisition, mobilization and recycling, and trigger a genome-wide metabolic remodeling of iron-dependent pathways. Here, we describe a mechanism that optimizes the activity of yeast ribonucleotide reductase when iron is scarce. We demonstrate that Aft1 and the DNA-binding protein Ixr1 enhance the expression of the gene encoding for its catalytic subunit, RNR1, in response to iron limitation, leading to an increase in both mRNA and protein levels. By mutagenesis of the Aft1-binding sites within RNR1 promoter, we conclude that RNR1 activation by iron depletion is important for Rnr1 protein and deoxyribonucleotide synthesis. Remarkably, Aft1 also activates the expression of IXR1 upon iron scarcity through an iron-responsive element located within its promoter. These results provide a novel mechanism for the direct activation of ribonucleotide reductase function by the iron-regulated Aft1 transcription factor.
真核生物核苷酸还原酶是一种依赖于铁的酶,可催化脱氧核苷酸从头合成的限速步骤。多种机制可响应遗传毒性应激和铁缺乏来调节核苷酸还原酶的活性。在缺铁时,酿酒酵母的 Aft1 转录因子特异性结合到一组称为铁调控基因的启动子内的铁反应顺式元件,激活其转录。铁调控基因的成员参与铁的摄取、动员和再循环,并引发与铁依赖途径相关的全基因组代谢重塑。在这里,我们描述了一种在铁缺乏时优化酵母核苷酸还原酶活性的机制。我们证明,Aft1 和 DNA 结合蛋白 Ixr1 响应铁限制增强编码其催化亚基 RNR1 的基因的表达,导致 mRNA 和蛋白质水平的增加。通过对 RNR1 启动子中 Aft1 结合位点的诱变,我们得出结论,铁耗竭时 RNR1 的激活对于 Rnr1 蛋白和脱氧核苷酸合成很重要。值得注意的是,Aft1 还通过位于其启动子内的铁反应元件在铁匮乏时激活 IXR1 的表达。这些结果为铁调控的 Aft1 转录因子直接激活核苷酸还原酶功能提供了一种新的机制。