The molecular basis for the induction of autophagy

诱导自噬的分子基础

• 批准号: 8199955
• 负责人: Michael Joseph Ragusa
• 金额: $ 4.52万
• 依托单位: U.S. NATIONAL INST DIABETES/DIGST/KIDNEY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8199955
• 关键词: Alkaline Phosphatase; Antineoplastic Agents; Architecture; Autophagocytosis; Binding; Biochemical; Biological Assay; Cancer Etiology; Catalytic Domain; Cell physiology; Cells; Cellular biology; Complex; Crystallization; Diabetes Mellitus; Disease; Docking; Drug Delivery Systems; Drug Design; Drug usage; Fractionation; Future; Genes; Goals; Homeostasis; Human; Huntington Disease; Hybrids; Image; Immune; Individual; Insecta; Lead; Lipids; Lysosomes; Malignant Neoplasms; Mammalian Cell; Measures; Mediating; Modeling; Molecular; Molecular Conformation; Molecular Sieve Chromatography; Molecular Weight; Monitor; Multiprotein Complexes; Neurodegenerative Disorders; Neurons; Organelles; Parkinson Disease; Pathway interactions; Pharmacologic Substance; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Process; Protein Biosynthesis; Protein Dephosphorylation; Protein Kinase; Proteins; Proteolysis; Regulation; Research; Saccharomyces cerevisiae; Sirolimus; Solutions; Specificity; Structure; Testing; Validation; Work; X-Ray Crystallography; Yeasts; base; design; in vitro Assay; in vivo; inhibition of autophagy; insight; interest; mutant; novel; overexpression; particle; protein complex; protein degradation; protein misfolding; reconstruction; research study; scaffold; therapeutic target; tumor progression

DESCRIPTION (provided by applicant): Protein turnover within a cell is regulated by protein synthesis and degradation. One pathway of protein degradation is autophagy, which is capable of degrading misfolded proteins and damaged organelles. A dysregulation of autophagy has been implicated in various cancers, and neurodegenerative diseases including Parkinson's and Huntington's disease. Due to the importance of autophagy in normal cellular function and its dysregulation in disease states there has been a significantly increased interest in autophagy related research within the last 15 years. Despite the increased interest in autophagy research, very few structures of autophagy related proteins have been determined. Therefore, many of the molecular mechanisms that regulate autophagy, including the mechanism of activation of the Atg1/ULK1 complex, remain to be elucidated. The Atg1/ULK1 is the most upstream complex that regulates autophagy and upon activation serves to initiate autophagy. Inhibition of Atg1/ULK1 complex activation results in the near complete inhibition of autophagy demonstrating that the Atg1/ULK1 complex is a master regulator of autophagy. Our goal is to understand the molecular mechanisms that regulate the initiation of autophagy by the Atg1/ULK1 complex, which will provide the molecular basis for the induction of autophagy. To this end, I will perform a structural and functional characterization of the inactive and active forms of the Atg1/ULK1 complex. This work will include structure determination of the Atg1/ULK1 complex as well as a structure driven mutational analysis of the complex to validate and further investigate possible mechanisms for complex activation. Additionally, autophagy has been shown to be induced by pharmaceutical drugs that are used to slow tumor progression. As a result, this work will not only reveal the molecular mechanisms for the induction of autophagy but may also drive future drug design towards targeting the Atg1/ULK1 complex to induce autophagy with higher specificity. PUBLIC HEALTH RELEVANCE: Autophagy, a process that helps maintain protein homeostasis, is dysregulated in numerous cancers and a number of pharmaceuticals that inhibit cancer progression act through the induction of autophagy. However, the mechanisms guiding the induction of autophagy are not understood. Therefore, gaining an understanding of these mechanisms will provide insight into the action of these cancer inhibiting pharmaceuticals, allow for the design of more potent cancer drugs, and additionally lead to novel drug targets.

描述（由申请人提供）：细胞内的蛋白质周转受蛋白质合成和降解的调节。蛋白质降解的一个途径是自噬，其能够降解错误折叠的蛋白质和受损的细胞器。自噬的失调与各种癌症和神经退行性疾病（包括帕金森病和亨廷顿病）有关。由于自噬在正常细胞功能中的重要性及其在疾病状态中的失调，在过去的15年中，对自噬相关研究的兴趣显着增加。尽管自噬研究的兴趣越来越大，但自噬相关蛋白的结构很少被确定。因此，许多调节自噬的分子机制，包括Atg 1/ULK 1复合物的激活机制，仍有待阐明。Atg 1/ULK 1是调节自噬的最上游复合物，并且在激活后用于启动自噬。Atg 1/ULK 1复合物激活的抑制导致自噬的几乎完全抑制，表明Atg 1/ULK 1复合物是自噬的主要调节剂。我们的目标是了解Atg 1/ULK 1复合物调控自噬启动的分子机制，这将为诱导自噬提供分子基础。为此，我将进行Atg 1/ULK 1复合物的非活性和活性形式的结构和功能表征。这项工作将包括Atg 1/ULK 1复合物的结构测定以及复合物的结构驱动突变分析，以验证和进一步研究复合物激活的可能机制。此外，自噬已被证明是由用于减缓肿瘤进展的药物诱导的。因此，这项工作不仅将揭示诱导自噬的分子机制，而且还可能推动未来的药物设计，以靶向Atg 1/ULK 1复合物，以更高的特异性诱导自噬。 公共卫生相关性：自噬是一种有助于维持蛋白质稳态的过程，在许多癌症中失调，并且许多抑制癌症进展的药物通过诱导自噬起作用。然而，指导自噬诱导的机制尚不清楚。因此，获得对这些机制的理解将提供对这些癌症抑制药物的作用的洞察，允许设计更有效的癌症药物，并另外导致新的药物靶点。