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：
7758262
负责人：
ROBERT CARL DICKSON
金额：
$ 2.12万
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 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7758262
关键词：
Affect Age Aging Alanine Alleles Animal Model Binding Biochemical Biogenesis Biological Assay Caloric Restriction Cell Fraction Cell Fractionation Cells Cellular Membrane Chimeric Proteins Chromatin Density Gradient Centrifugation Down-Regulation Enzymes Genes Genetic Genetic Screening Glass Homologous Gene Human In Vitro Ion-Exchange Chromatography Procedure Knowledge Label Libraries Longevity Longevity Pathway Mammals Mass Spectrum Analysis Measures Mediator of activation protein Membrane Molecular Mutate Organelles Oxidative Stress Pathway interactions Phase Phosphatidylinositols Phosphorylation Physiological Play Protein Binding Protein Biosynthesis Protein Kinase Proteins Proto-Oncogene Proteins c-akt Research Ribosomes Role Saccharomyces cerevisiae Serine Signal Transduction Pathway Slide Sphingolipids Temperature Testing Translation Initiation Work Yeasts alkyl hydroperoxide reductase analog base cell type dihydroceramide desaturase dosage in vivo mutant novel prevent public health relevance uptake yeast protein

项目摘要

DESCRIPTION (provided by applicant): Model organisms, such as Saccharomyces cerevisiae, have proven effective in elucidating genes and metabolites that modulate life span 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 phosphoinositide-dependent protein kinase 1 (PDK1), as the first upstream regulator 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 Ahp1 by Sch9 plays a role in CLS. We will also determine in vivo if LCBs and Pkh1 act upstream to regulate Sch9 during chronological aging. 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, newer automated but complementary strategies, and a genetic screening strategy. 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. PUBLIC HEALTH RELEVANCE: The proposed research will determine how the Sch9 protein kinase, a functional homolog of mammalian Akts/PKBs, regulates chronological life span in the Baker's yeast Saccharomyces cerevisiae. Signal transduction pathways that contain Akt/PKB type protein kinases play important roles in regulating aging and life span, and these pathways have been conserved from yeast to mammals. Thus, information gained from these studies should provide a more mechanistic basis for understanding how aging and life span are regulated in humans.

描述（由申请人提供）：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，并且这些知识构成了理解人类衰老和长寿的基础。这些研究已确定 Akt/PKB 蛋白激酶是保守的衰老调节因子。然而，尚不完全清楚它们是如何工作的。我们研究的重点之一是酿酒酵母 Sch9 蛋白激酶，它是一种调节时间寿命 (CLS) 的 AKT 同源物。通过体外研究，我们确定鞘脂长链碱基 (LCB) 和 Pkh1（人磷酸肌醇依赖性蛋白激酶 1 (PDK1) 的同源物）是 Sch9 的第一个上游调节因子。此外，我们还确定了第一个 Sch9 底物 Rli1（参与翻译起始和核糖体生物发生）和 Ahp1（一种防止氧化应激的烷基氢过氧化物还原酶）。我们将在这些新结果的基础上确定 Sch9 对 Rli1 和 Ahp1 的磷酸化是否在 CLS 中发挥作用。我们还将在体内确定 LCB 和 Pkh1 在时间老化过程中是否在上游发挥作用来调节 Sch9。尽管我们已经成功鉴定了 Sch9 底物，但可能还有其他底物，我们建议通过经典生化策略、更新的自动化但互补的策略以及遗传筛选策略来寻找新的底物。总之，这些研究的结果将为我们提供急需的关于酵母中 Akt (Sch9) 活性如何调节以及它如何通过下游底物调节 CLS 的机制理解。这些知识将有助于为理解人类衰老和长寿提供分子基础。公共健康相关性：拟议的研究将确定 Sch9 蛋白激酶（哺乳动物 Akts/PKB 的功能同源物）如何调节面包酵母酿酒酵母的时间寿命。含有Akt/PKB型蛋白激酶的信号转导通路在调节衰老和寿命方面发挥着重要作用，并且这些通路从酵母到哺乳动物都是保守的。因此，从这些研究中获得的信息应该为理解人类如何调节衰老和寿命提供更机械的基础。