DISTINCT REGULATION OF AUTOPHAGIC ACTIVITY BY TWO NOVEL PROTEINS

两种新型蛋白质对自噬活性的独特调节

• 批准号: 8169173
• 负责人: Zhenyu Yue
• 金额: $ 0.35万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169173
• 关键词: 1-Phosphatidylinositol 3-Kinase; Autophagocytosis; Autophagosome; Binding Proteins; Biochemical; Biochemistry; Cell Death; Cellular biology; Complex; Computer Retrieval of Information on Scientific Projects Database; Development; Endocytosis; Funding; Genetic; Grant; Institution; Lipids; Manuscripts; Mediating; Membrane; Mus; Nature; Nerve Degeneration; Pathway interactions; Phosphatidylinositols; Phosphotransferases; Play; Proteins; Publishing; Regulation; Research; Research Personnel; Resources; Role; Source; Structure; Tissues; Tumor Suppression; United States National Institutes of Health; Vacuole; Vesicle; Work; in vivo; novel; protein complex; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Beclin 1 is a mammalian autophagy protein which was shown to play an important role in development, tumor suppression, neurodegeneration and cell death. Beclin 1 forms a complex with Vps34/phosphatidylinositol (PtdIns) 3-kinase (class III PI-3K), which mediates multiple vesicle trafficking pathways including endocytosis and autophagy. However, the precise role of Beclin 1-Vps34/PtdIns3K complex in the regulation of autophagy remains to be elucidated. Through a study that combines mouse genetics and biochemistry, we uncover several protein components that associate specifically with Beclin 1 complexes in mouse tissues. The biochemical analysis reveals a large in vivo Beclin 1 complex containing the known proteins Vps34/PtdIns3K, p150/Vps15 and UVRAG, as well as novel proteins BISC and BIRC. Characterization of the novel proteins suggests that BISC and BIRC modulate Vps34/PtdIns3K activity in opposite manner. This opposing effect of BISC and BIRC on the lipid kinase activity is correlated with their distinct regulation of autophagy: BISC positively regulates autophagy, whereas BIRC is involved in negative control of autophagy. Moreover, we find that Beclin 1 and BISC synergistically promote formation of double membrane vacuoles which are associated with Atg5/Atg12, while forced expression of BIRC results in aberrant late endosomal/lysosomal structures (independent of Beclin 1) and impaired autophagosome maturation. Our study suggests that, by forming distinct protein complexes, Beclin 1 and its binding proteins orchestrate the precise function of Vps34/PtdIns3K in regulating autophagy at multiple steps. A manuscript describing this work has been published (18. Y. Zhong, Q.J. Wang, X. Li, B.T. Chait, N. Heintz, Z. Yue "Distinct regulation of autophagic activity by Atg14L and Rubicon associated with Beclin 1-phosphatidylinositol-3-kinase complex" Nature Cell Biology. 11 (2009) 468-76.)

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 Beclin 1是一种哺乳动物的自噬蛋白，被证明在发育，抑制，神经退行性和细胞死亡中起着重要作用。 Beclin 1与VPS34/磷脂酰肌醇（PTDINS）3-激酶（III PI-3K类）形成复合物，该酶介导了多个囊泡运输途径，包括内吞和自噬。但是，Beclin 1-VPS34/PTDINS3K复合物在自噬调节中的确切作用尚待阐明。通过将小鼠遗传学和生物化学结合的研究，我们发现了几种与小鼠组织中贝克林1络合物特别相关的蛋白质成分。生化分析揭示了一个大型体内Beclin 1复合物，其中含有已知的蛋白质VPS34/PTDINS3K，P150/VPS15和UVRAG，以及新型的蛋白质BISC和BICC。新型蛋白质的表征表明，BISC和BIRC以相反的方式调节VPS34/PTDINS3K活性。 BISC和BIRC对脂质激酶活性的这种相反的影响与它们的自动噬菌体的独特调节相关：BISC积极调节自噬，而BIRC则参与自噬的负面对照。此外，我们发现Beclin 1和BISC协同促进了与ATG5/ATG12相关的双膜液泡的形成，而强迫表达BIRC会导致晚期内体/溶酶体结构（与Beclin 1无关）和自噬体成熟。 我们的研究表明，通过形成不同的蛋白质复合物，Beclin 1及其结合蛋白在多个步骤中调节自噬在调节自噬中的VPS34/PTDINS3K的精确功能。一份描述这项工作的手稿已经发表（18。Y.Zhong，Q.J. Wang，X。Li，B.T。Chait，N。Heintz，Z。Yue，“ ATG14L对自噬活性的明显调节和与Beclin 1-磷酸脂蛋白肌醇 - 3-激酶复合物相关的Rubicon和Rubicon相关。