Dissecting the Molecular Mechanisms of Selective Autophagy

剖析选择性自噬的分子机制

• 批准号: 10004516
• 负责人: Michael Joseph Ragusa
• 金额: $ 40.5万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-14 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004516
• 关键词: Autophagocytosis; Autophagosome; Biochemistry; Biogenesis; Cellular biology; Complement; Complex; Event; Hybrids; Investigation; Knowledge; Laboratories; Lysosomes; Malignant Neoplasms; Membrane; Methods; Molecular; Nature; Nerve Degeneration; Organelles; Pathway interactions; Process; Proteins; Role; Structure; Surface; System; Vacuole; Vesicle; chronic infection; human disease; insight; pathogen; protein aggregation; protein function; reconstitution; structural biology; trafficking; yeast genetics

The accumulation of damaged organelles and large protein aggregates has been correlated with human diseases including cancer and neurodegeneration. Due to the large and complex nature of these cytosolic components, they must be degraded through a specialized vesicle trafficking pathway termed selective autophagy. In selective autophagy, large cytosolic material is captured within double membrane vesicles, termed autophagosomes, and targeted to the vacuole or lysosome for degradation. While the biogenesis of most trafficking vesicles occurs through the budding of a preexisting membrane surface, the biogenesis of the selective autophagosome occurs through a distinct process in which the cargo serves as a template for de novo vesicle biogenesis. The molecular mechanisms of these vesicle biogenesis events are largely unknown. The primary reason for this gap in our knowledge is that the structure and function of the proteins which function early in selective autophagy are unknown. In addition, many of these proteins lack obvious conserved domains which makes it challenging to predict a mechanism for these proteins. To investigate the structure and function of these proteins we will use hybrid structural biology methods, yeast genetics, biochemistry and cell biology. To complement our structure and function studies we will develop a method to reconstitute selective autophagy. This reconstitution system will ultimately enable us to evaluate what selective autophagy factors are required at each stage of vesicle biogenesis. Through these diverse investigations, we will establish a comprehensive description of the molecular mechanisms of selective autophagy. As selective autophagy has been increasingly correlated with human diseases, determining the mechanisms of selective autophagy will also provide invaluable insight into the complicated relationship between selective autophagy and human disease.

受损细胞器和大蛋白质聚集体的积累已被关联 与癌症和神经退行性疾病等人类疾病有关。由于规模庞大且复杂 由于这些细胞质成分的性质，它们必须通过专门的囊泡降解 贩运途径称为选择性自噬。在选择性自噬中，大的胞质物质 被捕获在双膜囊泡内，称为自噬体，并靶向 液泡或溶酶体进行降解。虽然大多数运输囊泡的生物发生发生 通过预先存在的膜表面的出芽，选择性的生物发生 自噬体通过一个独特的过程发生，其中货物作为模板 从头囊泡生物发生。这些囊泡生物发生事件的分子机制是 很大程度上不为人知。我们的知识差距的主要原因是结构和 在选择性自噬早期起作用的蛋白质的功能尚不清楚。此外， 许多这些蛋白质缺乏明显的保守结构域，这使得预测 这些蛋白质的作用机制。为了研究这些蛋白质的结构和功能，我们将 使用混合结构生物学方法、酵母遗传学、生物化学和细胞生物学。到 补充我们的结构和功能研究，我们将开发一种重组方法 选择性自噬。这个重组系统最终将使我们能够评估什么 囊泡生物发生的每个阶段都需要选择性自噬因子。通过这些 通过多样化的研究，我们将建立对分子的全面描述 选择性自噬机制。随着选择性自噬的相关性越来越大 对于人类疾病，确定选择性自噬机制也将提供 对选择性自噬与人类之间复杂关系的宝贵见解 疾病。