The mechanism and regulation of autophagy

自噬代表了一种复杂的细胞内稳态途径，起到了细胞保护作用，但如果 调控失调可导致细胞死亡；对调控的全面了解对潜在的调控至关重要 这一过程用于治疗目的，并增加我们对膜动力学和 细胞器生物发生。自噬发生在所有真核生物中，自噬的蛋白质成分 从酵母到哺乳动物，机械都是保守的。这一过程的特点是形成了双重的-- 细胞膜液泡，自噬小体，含有细胞质成分，由 吞噬动物，一种独特的瞬变隔间。完成后，自噬小体与 溶酶体/液泡释放被分解的内部小泡，允许接近货物。自噬 在不同的发育过程中发挥作用，并与一系列病理生理 条件。这项建议的长期目标是了解自噬的作用机制- 相关蛋白质，这一过程是如何调控的，以及为什么特定的突变会导致人类疾病。 在过去的20年里，我们对自噬-42自噬的分子方面了解了很多- 相关(ATG)蛋白已经被鉴定，但我们对它们的功能只有一个粗略的了解。此外, 自噬功能障碍与人类的许多疾病有关，包括癌症、心脏病和 神经退行性变。虽然自噬主要是细胞保护，但过度的自噬是有害的。因此， 我们需要对监管网络有充分的了解，以便应用旨在 通过自噬调节来治疗疾病。在接下来的五年里，我们希望(1)了解结构 和ATG蛋白的功能；(2)定义允许自噬诱导的调控控制，确定 在特定和非特定类型的自噬之间切换，并在适当的时候保持自噬 水平；以及(3)破译影响自噬的突变和疾病之间的关系， 促进治疗的最终目标。我们正在使用酵母来研究它的分子机制 自噬：这是对这一复杂过程进行分子遗传和生化分析的最佳系统。 然而，由于高度的保守性，我们从酵母中学到的信息将适用于 到更复杂的真核生物。 这项提案中描述的实验具有重要意义，因为它们将阐明重要的联系 在上游调控组件和执行自噬的机器之间，提供下一个 在将信号转导元件链接到功能设备的综合分析中的步骤， 提高我们的基础细胞生物学知识，并确定最终治疗干预的目标。 拟议的研究是创新的，因为它提供了新的，在某些情况下是范式转移， 关于自噬的调节和功能组件的信息。

Autophagy represents a complex pathway of cellular homeostasis that functions in cytoprotection, but if dysregulated can cause cell death; a complete knowledge of regulation is critical for the potential modulation of this process for therapeutic purposes, and to increase our basic understanding of membrane dynamics and organelle biogenesis. Autophagy occurs in all eukaryotes, and the protein components of the autophagic machinery are conserved from yeast to mammals. The hallmark of this process is the formation of double- membrane cytosolic vesicles, autophagosomes, that contain cytoplasmic components sequestered by the phagophore, a unique transient compartment. After completion, the autophagosomes fuse with the lysosome/vacuole to release the inner vesicle that is broken down, allowing access to the cargo. Autophagy plays a role in various developmental processes and is associated with a range of pathophysiological conditions. The long-term goals of this proposal are to understand the mechanism of action of the autophagy- related proteins, how the process is regulated, and why specific mutations result in human disease. In the last 20 years, we have learned much about the molecular aspects of autophagy—42 autophagy- related (Atg) proteins have been identified, but we only have a cursory knowledge of their function. In addition, autophagic dysfunction is associated with numerous diseases in humans, including cancer, heart disease and neurodegeneration. Although autophagy is primarily cytoprotective, excessive autophagy is detrimental. Thus, we need a full understanding of the regulatory network in order to apply therapeutic interventions aimed at treating disease through autophagy modulation. In the next five years, we want to (1) understand the structure and function of the Atg proteins; (2) define the regulatory controls that allow autophagy induction, determine the switch between specific and non-specific types of autophagy, and maintain autophagy at appropriate levels; and (3) decipher the relationship between mutations affecting autophagy and diseases, with an ultimate goal of facilitating treatment. We are using yeast to investigate the molecular mechanism of autophagy; this is the best system for a molecular genetic and biochemical analysis of this complex process. Because of the high degree of conservation, however, the information we learn from yeast will be applicable to more complex eukaryotes. The experiments described in this proposal are significant because they will elucidate important links between upstream regulatory components and the machinery that carries out autophagy, providing the next step in a comprehensive analysis that links the signal transduction elements to the functional apparatus, advancing our knowledge of basic cell biology, and identifying targets for ultimate therapeutic intervention. The proposed research is innovative, because it is providing new, and in some cases paradigm-shifting, information about the regulatory and functional components of autophagy.