Biogenesis of transport vesicles coated by COPI

COPI 包被的运输囊泡的生物发生

• 批准号: 7807393
• 负责人: VICTOR W HSU
• 金额: $ 48.92万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7807393
• 关键词: ADP-Ribosylation Factors; ARFGAP1; Affect; Binding; Biochemical; Biogenesis; Capsid Proteins; Cell membrane; Cells; Clathrin; Coat Protein Complex I; Complex; Coupled; Ear; Endocytosis; Event; Funding; GTPase-Activating Proteins; Goals; Golgi Apparatus; Guanine Nucleotide Exchange Factors; Image; In Transferrin; Investigation; Iron; Maps; Mediating; Membrane Proteins; Molecular; Monomeric GTP-Binding Proteins; Mutation; Parents; Pathologic; Pathway interactions; Point Mutation; Process; Proteins; Recovery; Regulation; Role; Sorting - Cell Movement; Transcription Factor AP-2 Alpha; Transferrin Receptor; Transport Vesicles; United States National Institutes of Health; Vesicle; insight; iron metabolism; novel; parent grant; protein function; public health relevance; response; uptake; ADP-Ribosylation Factors; ARFGAP1; Affect; Binding; Biochemical; Biogenesis; Capsid Proteins; Cell membrane; Cells; Clathrin; Coat Protein Complex I; Complex; Coupled; Ear; Endocytosis; Event; Funding; GTPase-Activating Proteins; Goals; Golgi Apparatus; Guanine Nucleotide Exchange Factors; Image; In Transferrin; Investigation; Iron; Maps; Mediating; Membrane Proteins; Molecular; Monomeric GTP-Binding Proteins; Mutation; Parents; Pathologic; Pathway interactions; Point Mutation; Process; Proteins; Recovery; Regulation; Role; Sorting - Cell Movement; Transcription Factor AP-2 Alpha; Transferrin Receptor; Transport Vesicles; United States National Institutes of Health; Vesicle; insight; iron metabolism; novel; parent grant; protein function; public health relevance; response; uptake

DESCRIPTION (provided by applicant): We are submitting a revision to the parent R01 application (GM058615) in response to an NIH announcement (NOT-OD-09-058: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications). The parent grant studies transport vesicles formed by the Coat Protein I (COPI) complex. This revision studies a new topic outside the scope of the parent grant, examining how a key COPI component, known as ARFGAP1, has a novel role in clathrin-mediated endocytosis. The clathrin AP2 complex participates in endocytosis from the plasma membrane, while COPI acts in transport from the Golgi to the ER and also among the Golgi stacks. These two coat complexes are the first ones identified, and have not been shown to share a common component despite having been intensely investigated for many years. We have now gathered evidence that ARFGAP1 acts in a subset of clathrin AP2-dependent transport, as defined by the endocytosis of transferrin receptor (TfR), and propose to further elucidate this process through two major approaches. First, using biochemical approaches, we will examine whether the interaction between AP2 and ARFGAP1 regulates the ability of either component in binding to TfR. We also seek insight into how ARFGAP1 interacts with AP2 by defining a minimal portion of ARFGAP1 that binds to the 1-ear domain, and also map how ARFGAP1 binds to the 1-ear domain. Second, we will pursue advanced imaging approaches to interrogate how the GAP activity affects vesicle formation and cargo sorting, and whether the interaction between ARFGAP1 and AP2 also affect these two events. We anticipate that potential results will contribute to a general understanding of mechanisms in vesicular transport. Moreover, because TfR endocytosis is essential for iron uptake, anticipated results will also contribute to a molecular understanding of how iron uptake is achieved, and how this process may become pathologic. PUBLIC HEALTH RELEVANCE: The function of proteins is critically regulated by their localization. This localization is achieved in part by transport pathways that act as highways within the cell. We propose to understand how a regulator of this process modulates the distribution of a surface protein known as the transferrin receptor. Our results will likely contribute to a basic understanding of transport mechanisms within the cell, and also shed insight into the regulation of iron metabolism.

描述（由申请人提供）：我们正在向父母R01申请（GM058615）提交修订，以响应NIH公告（NOT-OD-09-058：NIH宣布竞争性修订申请的恢复ACT资金的可用性）。父授予研究由外套I（COPI）复合物形成的传输囊泡。这项修订研究了父母赠款范围之外的一个新主题，研究了一个称为ArfGAP1的关键COPI组件如何在网格蛋白介导的内吞作用中具有新颖的作用。网格蛋白AP2复合物参与了质膜内吞作用，而COPI起着从高尔基人到ER以及高尔基体堆栈中的运输。这两个外套配合物是第一个鉴定的配合物，尽管经过多年进行了深入研究，但尚未显示出共同的组成部分。现在，我们收集的证据表明，ARFGAP1在网状蛋白AP2依赖性转运的一部分中作用，这是由转运蛋白受体（TFR）的内吞作用所定义的，并建议通过两种主要方法进一步阐明这一过程。首先，使用生化方法，我们将检查AP2和ARFGAP1之间的相互作用是否调节任何两分组成在与TFR结合中的能力。我们还寻求了解ARFGAP1如何通过定义与1-EAR结构域结合的ARFGAP1的最小部分与AP2相互作用的信息，并映射ARFGAP1如何与1-EAR域结合。其次，我们将采用先进的成像方法来询问间隙活动如何影响囊泡的形成和货物排序，以及ARFGAP1和AP2之间的相互作用是否也影响了这两个事件。我们预计潜在的结果将有助于对囊泡运输机制的一般理解。此外，由于TFR内吞作用对于铁的摄取至关重要，因此预期的结果也将有助于分子了解如何实现铁的摄取，以及该过程如何变得病理性。 公共卫生相关性：蛋白质的功能受其本地化的严格调节。这种定位是通过在电池内充当高速公路的运输途径来实现的。我们建议了解该过程的调节剂如何调节称为转铁蛋白受体的表面蛋白的分布。我们的结果可能会有助于对细胞内的运输机制的基本理解，并深入了解铁代谢的调节。