Sir2 regulation and chemical modulation

拟议研究的目标是推导出控制 酵母和人体细胞中sirtuin酶的活性。这些原则将被用来设计小型 可以激活细胞中sirtuins的分子。Sirtuins调节一系列具有生物学意义的活动。 包括抗逆性、基因沉默和长寿。Sirtuins参与调节生物 与卡路里限制相关的影响。卡路里限制已被证明有许多积极的作用 对哺乳动物的健康益处包括减少脂肪生成、胰岛素敏感性和延长寿命。 这些生物学效应引起了人们对sirtuins的酶机制的兴趣，sirtuins通过这种方式 它们是在细胞中被调节的，以及它们可能被药理学调节以改进的方式 人类健康。Sirtuins是一种依赖NAD的脱乙酰基酶，可以去除修饰的乙酰赖氨酸的乙酰基 组蛋白和转录因子，从而调节染色质和基因表达。我们和其他人有 证明这些活动受细胞内NAD和烟酰胺水平的调节，并且 Sirtuins能够整合来自能量和新陈代谢状态的信息来控制遗传事件。作为一部分 我们的目标是更好地了解这些酶的功能以及它们如何在细胞中被调节 建议研究以下具体目标：在Aim1中，我们建议表征生物化学 这类独特的酶的功能，强调它们的化学新颖性和NAD的结合 在脱乙酰化反应中。在AIM2中，我们建议展示酶活性如何提供一种机制 烟酰胺对sirtuin活性的调节。此外，最近开发的一种质谱学方法 我们希望对烟酰胺如何调节细胞中的sirtuins获得新的见解。在目标3中，我们探索 根据sirtuin反应知识设计sirtuins小分子激活剂的研究进展 烟酰胺的调节机制和作用机制。这些激活剂体现了一种新的方法 上调细胞中sirtuin的作用，为药物干预提供一个潜在的切入点 提高细胞的抗应激能力和细胞存活率。这些目标的实现有望提供新的 对这些酶的生物化学和调节的洞察，并为新的 可以激活sirtuins来治疗糖尿病和退行性疾病的治疗方法。

The objective of the proposed research is to deduce mechanistic and regulatory principles that control the activities of sirtuin enzymes in yeast and human cells. These principles will be used to design small molecules that can activate sirtuins in cells. Sirtuins regulate a host of biologically significant activities including stress resistance, gene silencing and longevity. Sirtuins are implicated in mediating biological effects associated with calorie restriction. Calorie restriction has been shown to have numerous positive health benefits in mammals including reduced adipogenesis, insulin sensitivity and increased lifespan. These biological effects have raised interest in the enzymatic mechanisms of sirtuins, the means by which they are regulated in cells and the ways in which they might be modulated pharmacologically for improved human health. Sirtuins are NAD dependent deacetylases that remove acetyl-groups of acetyllysine modified histones and transcription factors thereby regulating chromatin and gene expression. We and others have demonstrated that these actvitities are regulated by NAD and nicotinamide levels in cells, and that the sirtuins are able to integrate information from energy and metabolic states to control genetic events. As part of our goal to better understand the functions of these enzymes and how they can be modulated in cells we propose to investigate the following specific aims: In Aim1 we propose to characterize the biochemical functions of this unique class of enzymes, emphasizing their chemical novelty and the incorporation of NAD in deacetylation reactions. In Aim2 we propose to show how enzymatic activity provides a mechanism for nicotinamide regulation of sirtuin activity. In addition with a recently developed mass spectrometry method we hope to gain new insights into how nicotinamide regulates sirtuins in cells. In Aim 3 we explore the development of small molecule activators of sirtuins designed from knowledge of the sirtuin reaction mechanism and the mechanism of nicotinamide regulation. These activators embody a novel approach to upregulate sirtuin action in cells and provide a potential entrypoint for pharmacological intervention to increase cell stress resistance and cell survival. Achievement of these aims is expected to provide new insights into the biochemistry and regulation of these enzymes, and provide proof of concept for new therapeutics that can activate sirtuins to treat diabetes and degenerative disorders.