Characterization of the Sch9 -Longevity Pathway in Yeast

酵母中 Sch9 长寿途径的表征

基本信息

批准号：
7671879
负责人：
ROBERT CARL DICKSON
金额：
$ 36.63万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-01 至 2008-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7671879
关键词：
Aging Alanine Alleles Animal Model Binding Biochemical Biogenesis Biological Assay Caloric Restriction Cell Fractionation Cells Cellular Membrane Chimeric Proteins Chromatin Data Density Gradient Centrifugation Enzymes Genes Glass Homologous Gene Human In Vitro Ion-Exchange Chromatography Procedure Knowledge Label Libraries Longevity Longevity Pathway Mass Spectrum Analysis Measures Mediating Mediator of activation protein Molecular Mutate Organelles Oxidative Stress Pathway interactions Phase Phosphatidylinositols Phosphorylation Phosphorylation Site Physiological Play Protein Binding Protein Biosynthesis Protein Kinase Proteins Proto-Oncogene Proteins c-akt Publishing RAC-Alpha Serine/Threonine Kinase Radiolabeled Rate Research Ribosomes Role Role playing therapy Saccharomyces cerevisiae Site Slide Sphingolipids Temperature Testing Translation Initiation Work Yeasts abstracting alkyl hydroperoxide reductase analog base cell type in vivo mutant novel prevent radiotracer yeast protein

项目摘要

Project Summary/Abstract Model organisms, such as Saccharomyces cerevisiae, have proven effective in elucidating genes and metabolites that modulate lifespan and this knowledge forms a basis for understanding human aging and longevity. These studies have identified the Akt/PKB protein kinases as conserved regulators of aging. However, it is not entirely clear how they work. One focus of our research is the S. cerevisiae Sch9 protein kinase, an AKT homolog that regulates chronological life span (CLS). From in vitro studies we identified sphingolipid long chain bases (LCBs) and Pkh1, a homolog of human phosphoinositidedependent protein kinase 1 (PDK1), as the first upstream regulators of Sch9. In addition, we identified the first Sch9 substrates, Rli1, involved in translation initiation and ribosome biogenesis, and Ahp1, an alkyl hydroperoxide reductase that protects against oxidative stress. We will build upon these novel results and determine if phosphorylation of Rli1 and Ahp1by Sch9 plays a role in CLS. We will also determine in vivo if LCBs and Pkh1 act upstream to regulate Sch9 during chronological aging. Published data suggest that the TOR proteins also regulate Sch9 during aging and we will determine if this hypothesis is true. Although we have successfully identified Sch9 substrates, there are likely to be others and we propose to search for new substrates by classical biochemical strategies and newer automated but complementary strategies. In summary, the results of these studies will provide the much needed mechanistic understanding of how Akt (Sch9) activity is regulated in yeast and how it regulates CLS through downstream substrates. This knowledge will help to provide a molecular basis for understanding human aging and longevity.

项目概要/摘要模型生物，例如酿酒酵母，已被证明可以有效地阐明调节寿命的基因和代谢物，这些知识构成了基础了解人类衰老和长寿。这些研究已确定 Akt/PKB 蛋白激酶作为衰老的保守调节剂。然而，尚不完全清楚如何他们工作。我们研究的一个重点是酿酒酵母 Sch9 蛋白激酶，一种 AKT 调节时间寿命（CLS）的同源物。从体外研究中我们发现鞘脂长链碱基 (LCB) 和 Pkh1（人磷酸肌醇依赖性蛋白的同源物）蛋白激酶1 (PDK1)，作为Sch9的第一个上游调节因子。此外，我们鉴定了第一个 Sch9 底物 Rli1，参与翻译起始和核糖体生物发生和 Ahp1，一种烷基氢过氧化物还原酶，可防止氧化压力。我们将在这些新结果的基础上确定 Rli1 和 Rli1 的磷酸化是否 Ahp1by Sch9 在 CLS 中发挥作用。我们还将确定 LCB 和 Pkh1 是否在体内发挥作用上游在时间老化过程中调节 Sch9。公布的数据表明 TOR 蛋白也在衰老过程中调节 Sch9，我们将确定这个假设是否成立真的。虽然我们已经成功鉴定了 Sch9 底物，但很可能其他人，我们建议通过经典生化策略寻找新的底物以及更新的自动化但互补的策略。综上所述，这些结果研究将为 Akt (Sch9) 活性提供急需的机制理解在酵母中受到调节，以及它如何通过下游底物调节 CLS。这知识将有助于为理解人类衰老和长寿。