Functional genomic approach to identify novel genes involved in the regulation of oxidative stress resistance and animal lifespan
Functional genomic approach to identify novel genes involved in the regulation of oxidative stress resistance and animal lifespan
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DOI：
10.1111/j.1474-9726.2007.00302.x
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发表时间：
2007-08-01
期刊：
影响因子：
7.8
通讯作者：
Sun, Hong
中科院分区：
文献类型：
--
作者：
Kim, Yongsoon;Sun, Hong
许多生物的遗传学研究表明，动物寿命延长的表型往往伴随着对活性氧(ROS)的抵抗力增强。在秀丽线虫中，编码胰岛素/胰岛素样生长因子1受体样分子的daf-2突变会导致动物寿命延长，并增加对ROS的抵抗力。我们已经优化了一种使用产生ROS的化学物质百草枯来监测蠕虫对ROS抗性的方法。我们已经使用这种方法来筛选染色体III和IV上的RNAi文库，寻找当沉默时授予百草枯抗性的基因。随后对阳性RNAi克隆进行了寿命延长表型筛选。使用这种方法，我们已经确定了84个基因，当被RNAi灭活时，会导致动物寿命的显著增加。在84个基因中，发现29个基因的作用方式依赖于daf-16。DAF-16是一种叉头转录因子，已知可以整合来自多个途径的信号，包括daf-2途径，以调节动物的寿命。84个基因中的大多数以前没有与衰老相关，它们可能参与重要的细胞过程，如信号转导、细胞-细胞相互作用、基因表达、蛋白质降解和能量代谢。我们的筛查还发现了一组基因，这些基因可能在调节线虫寿命的营养感应途径中发挥作用。我们的研究提供了一种新的方法来识别与衰老调节有关的基因。
Genetic studies in many organisms suggest that an increased animal lifespan phenotype is often accompanied by enhanced resistance toward reactive oxygen species (ROS). In Caenorhabditis elegans, mutations in daf-2, which encode an insulin/insulin-like growth factor 1 receptor-like molecule, lead to an extended animal lifespan and increased resistance to ROS. We have optimized an assay to monitor ROS resistance in worms using the ROS-generating chemical paraquat. We have employed this assay to screen the RNAi library along chromosomes III and IV for genes that, when silenced, confer paraquat resistance. The positive RNAi clones were subsequently screened for a lifespan extension phenotype. Using this approach, we have identified 84 genes that, when inactivated by RNAi, lead to significant increases in animal lifespan. Among the 84 genes, 29 were found to act in a manner dependent on daf-16. DAF-16, a forkhead transcription factor, is known to integrate signals from multiple pathways, including the daf-2 pathway, to regulate animal lifespan. Most of the 84 genes have not been previously linked to aging, and potentially participate in important cellular processes such as signal transduction, cell-cell interaction, gene expression, protein degradation, and energy metabolism. Our screen has also identified a group of genes that potentially function in a nutrient-sensing pathway to regulate lifespan in C. elegans. Our study provides a novel approach to identify genes involved in the regulation of aging.