The regulation of autophagy pathways in eukaryotic cells

真核细胞自噬途径的调控

• 批准号: 8309101
• 负责人: Paul K Herman
• 金额: $ 30.88万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8309101
• 关键词: Autophagocytosis; Biological; Biology; Cell Survival; Clinical; Complex; Crohn' s disease; Cyclic AMP-Dependent Protein Kinases; Disease; Drug Delivery Systems; Enzymes; Eukaryota; Eukaryotic Cell; Event; Genetic; Goals; Growth; Health; Human; Huntington Disease; Light; Link; Lipids; Malignant neoplasm of ovary; Mammals; Mediating; Membrane Protein Traffic; Metabolism; Microscopy; Modification; Molecular; Molecular Biology; Natural Immunity; Organelles; Organism; Pathway interactions; Phosphatidylinositols; Phosphorylation; Phosphotransferases; Physiological; Process; Protein Kinase; Proteins; Regulation; Rest; Role; Saccharomyces cerevisiae; Signal Pathway; Signal Transduction; Testing; Therapeutic; Therapeutic Intervention; Tumor Suppression; Work; Yeasts; drug discovery; enzyme substrate; insight; interest; malignant breast neoplasm; nervous system disorder; novel; phosphatidylinositol 3,5-diphosphate; protein complex; research study; stem; therapeutic target

DESCRIPTION (provided by applicant): We are interested in developing a better understanding of the biology of the non-dividing resting states that eukaryotic cells enter when conditions are not conducive to continued growth. One of our primary goals is to define how the processes that are induced during these periods of quiescence are regulated and how they contribute to general cell survival. This proposal is focused upon a set of related pathways, known collectively as autophagy, that are required for this survival. Autophagy pathways are responsible for the turnover of cytoplasmic material, including bulk protein and damaged or superfluous organelles. This autophagy-mediated degradation has been linked to a variety of processes relevant to human health, including tumor suppression, innate immunity and neurological disorders, such as Huntington's disease. In many of these conditions, the autophagy pathway is being considered as a major point of therapeutic intervention. It is therefore critical that we develop a thorough understanding of the mechanisms normally controlling autophagy in eukaryotic cells. This proposal will examine two key aspects of the regulation of autophagy in the yeast, Saccharomyces cerevisiae. Studies with this organism have contributed tremendously to our basic understanding of autophagy in all eukaryotes, including humans. First, a combination of approaches will be used to characterize the autophagy process induced by the inactivation of the cAMP-dependent protein kinase (PKA) signaling pathway. Our preliminary work indicates that this PKA-regulated process may be similar to an alternative form of macroautophagy recently identified in mammals. The experiments here will explore this possibility and define the molecular machineries governing this PKA-regulated pathway. These studies will also characterize a potential role for the phosphoinositide, PtdIns (3,5)P2, in this PKA-regulated macroautophagy. The second major goal of this proposal is the identification of the substrates of the Atg1 protein kinase. Atg1 is a key regulatory target within the autophagy machinery and identifying the targets of this enzyme represents one of the major goals in the autophagy field today. In all, we feel that the completion of this work will provide important insights into the manner in which autophagy is controlled in eukaryotic cells, insights that should facilitate efforts to manipulate this pathway in clinically beneficial ways. The specific aims in this proposal are: (1) to characterize the autophagy pathway induced upon inactivation of PKA signaling; (2) to characterize the link between PKA signaling, phosphoinositide metabolism and the regulation of autophagy; and (3) to identify and characterize substrates of the Atg1 protein kinase.

描述（由申请人提供）：我们有兴趣更好地了解非分散静止状态的生物学，即在条件不利于持续生长的情况下，真核细胞进入。我们的主要目标之一是定义如何调节这些静止期间诱导的过程以及它们如何促进一般细胞存活。该建议集中在此生存所需的一系列相关途径上，统称为自噬。自噬途径负责细胞质材料的营业额，包括块状蛋白质和受损或多余的细胞器。这种自噬介导的降解与与人类健康相关的各种过程有关，包括肿瘤抑制，先天免疫和神经系统疾病，例如亨廷顿氏病。在许多情况下，自噬途径被视为治疗干预的主要点。因此，至关重要的是，我们对通常控制真核细胞自噬的机制有透彻的理解。 该提案将研究酵母中自噬的两个关键方面，即酿酒酵母。对这种生物体的研究为我们在包括人类在内的所有真核生物中对自噬的基本理解做出了巨大贡献。首先，将使用多种方法来表征依赖CAMP依赖性蛋白激酶（PKA）信号通路引起的自噬过程。我们的初步工作表明，该PKA调节的过程可能类似于最近在哺乳动物中发现的大型噬菌体的替代形式。这里的实验将探讨这种可能性，并定义管理PKA调节途径的分子机器。这些研究还将表征在这种受PKA调节的大噬细胞中磷酸肌醇PTDINS（3,5）P2的潜在作用。该提案的第二个主要目标是鉴定ATG1蛋白激酶的底物。 ATG1是自噬机械内的关键调节目标，并确定该酶的目标是当今自噬领域的主要目标之一。总的来说，我们认为这项工作的完成将为真核细胞中自动噬的方式提供重要的见解，这些见解应促进以临床上有益的方式努力操纵这一途径。该提案中的具体目的是：（1）表征PKA信号失活而引起的自噬途径； （2）表征PKA信号传导，磷酸肌醇代谢和自噬调节之间的联系； （3）识别和表征ATG1蛋白激酶的底物。