Regulation of lifespan

寿命调节

基本信息

批准号：
9926773
负责人：
Kurt W Runge
金额：
$ 32.49万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-15 至 2022-04-30
项目状态：
已结题

项目摘要

Project Summary The aging population and coincident rise in associated diseases demand a better understanding of the basic mechanisms of aging to design appropriate interventions and prolong human health. Elucidating the molecular mechanisms of the processes that slow aging, such as lifespan extension by caloric or dietary restriction, would provide prime candidates for therapeutic intervention. Model organisms have allowed major advances in defining aging mechanisms due to their exceptionally powerful genetics and analytical tools, and because these processes are evolutionarily conserved from humans to yeasts. We discovered a new pathway in Schizosaccharomyces pombe that both regulates autophagy and extends lifespan. Autophagy is a process that degrades and recycles proteins and organelles, and has been linked to lifespan and diseases associated with human aging. We found that both autophagy and lifespan are regulated through the cyclin-dependent kinase Pef1, an ortholog of human Cdk5. Pef1 acts with its cyclin Clg1 to limit lifespan and autophagy, and is opposed by the effector kinase Cek1 that extends lifespan and increases autophagy levels. The Clg1-Pef1- Cek1 pathway acts independently of TOR, the only other pathway known to regulate both lifespan and autophagy, but how these processes are controlled by the Pef1 pathway is unknown. Aim 1 will test the hypothesis that the Pef1 pathway senses nutrients to control lifespan and autophagy. This aim will be significant for testing the linkage between autophagy and lifespan extension by caloric or dietary restriction. Establishing the mechanism by which the Pef1 pathway regulates lifespan and autophagy requires knowing the downstream effectors, and Aim 2 describes a novel chemical genomics approach to identify Pef1 targets. We modified the Pef1 kinase to allow selective binding of ATP analogs that tag kinase substrates to allow their isolation, and used the Pef1 variant to identify the substrates. We identified substrates with predicted roles in autophagy and lifespan, as well as substrates that we hypothesize mediate Pef1 control through signaling pathways. Aim 2 will validate the substrates and identify substrates of the effector kinase Cek1. Aim 3 will determine how the phosphorylation of Pef1 and Cek1 substrates affects specific processes in lifespan and autophagy control, and will also use high-throughput functional characterization of substrates to reveal signaling pathways regulated by the Pef1 pathway. The strong functional homology between Pef1 and human Cdk5 indicates that the Clg1-Pef1-Cek1 pathway, its substrates and functions will be conserved in humans, which we will also test in Aim 3. The results of these proposed studies will provide a mechanistic framework for a new, conserved lifespan and autophagy-regulating pathway with important ramifications for healthy human aging.

项目摘要相关疾病的人口老龄化和巧合的增长要求更好地了解基本衰老的机制设计适当的干预措施并延长人类健康。阐明分子缓慢衰老的过程的机制，例如通过热量或饮食限制延长寿命，将为治疗干预提供主要候选人。模型有机体允许重大进步由于其异常强大的遗传学和分析工具，定义衰老机制，因为这些过程从人类到酵母都是进化保守的。我们在调节自噬并延长寿命的精神分裂症POMBE。自噬是一个过程降解并回收蛋白质和细胞器，并与寿命和疾病有关随着人类的衰老。我们发现自噬和寿命都通过细胞周期蛋白依赖性调节激酶PEF1，人类CDK5的直系同源物。 PEF1用其细胞周期蛋白CLG1起作用以限制寿命和自噬，并且是与效应子激酶CEK1相反，它延长了寿命并增加了自噬水平。 clg1-pef1- CEK1途径独立于TOR，这是唯一已知的寿命和寿命调节的途径自噬，但是这些过程如何由PEF1途径控制。 AIM 1将测试假设PEF1途径可以感觉到营养能够控制寿命和自噬。这个目标将是对于通过热量或饮食限制来测试自噬和寿命扩展之间的连接意义很大。建立PEF1途径调节寿命和自噬的机制需要知道下游效应子，AIM 2描述了一种新型的化学基因组学方法来识别PEF1靶标。我们修改了PEF1激酶，以允许对ATP类似物进行选择性结合，该ATP类似物标记激酶底物以允许其隔离，并使用PEF1变体识别底物。我们确定了在自噬和寿命，以及我们通过信号来假设介导PEF1控制的底物途径。 AIM 2将验证底物并识别效应激酶CEK1的底物。目标3意志确定PEF1和CEK1底物的磷酸化如何影响生命周期中的特定过程自噬控制由PEF1途径调节的信号通路。 PEF1与人之间的强大功能同源 CDK5表明CLG1-PEF1-CEK1途径，其底物和功能将在人类中保存，我们还将在AIM 3中进行测试。这些提出的研究的结果将提供机械框架对于新的，保守的寿命和自噬调节途径，并具有重要的后果人类衰老。