Regulation of lifespan

项目摘要 人口老龄化和相关疾病的增加要求更好地理解基本的 老龄化机制，以设计适当的干预措施，延长人类健康。阐明分子 延缓衰老的机制，如通过限制卡路里或饮食来延长寿命， 将为治疗干预提供最佳候选者。模式生物使重大进展成为可能 在定义衰老机制方面，由于他们异常强大的遗传学和分析工具，以及因为 从人类到酵母，这些过程在进化上是保守的。我们发现了一条新的路径 裂殖酵母既能调节自噬又能延长寿命。自噬是一个过程 这会降解和回收蛋白质和细胞器，并与寿命和相关疾病有关 随着人类年龄的增长。我们发现，自噬和寿命都是通过依赖于细胞周期蛋白的 激酶Pef1，人CDK5的同源基因。Pef1与其周期蛋白Clg1一起作用于限制寿命和自噬，并且是 与效应蛋白激酶CEK1相反，CEK1可延长寿命并增加自噬水平。Clg1-Pef1- CEK1途径的作用独立于TOR，TOR是已知的唯一同时调节寿命和 自噬，但这些过程是如何由Pef1途径控制的还不清楚。目标1将测试 假设Pef1通路感知营养物质来控制寿命和自噬。这一目标将是 对于测试自噬和通过限制卡路里或饮食延长寿命之间的联系具有重要意义。 建立Pef1通路调节寿命和自噬的机制需要知道 和Aim 2描述了一种新的化学基因组学方法来识别Pef1靶标。 我们对Pef1激酶进行了修饰，以允许标记激酶底物的ATP类似物选择性结合，从而允许它们的 分离，并用Pef1变异体鉴定底物。我们确定了具有预测作用的底物 自噬和寿命，以及我们假设通过信号调节Pef1控制的底物 小路。AIM 2将验证底物并鉴定效应蛋白激酶CEK1的底物。目标3将 确定Pef1和CEK1底物的磷酸化如何影响寿命和 自噬控制，并将利用高通量的底物功能表征来揭示 受Pef1通路调控的信号通路。Pef1与人类的强功能同源性 CDK5表明Clg1-Pef1-CEK1途径及其底物和功能在人类中将是保守的， 我们还将在目标3中进行测试。这些拟议研究的结果将提供一个机制框架 一种新的、保守的寿命和自噬调节途径，对健康具有重要影响 人类的衰老。

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.