Protein Interactions Controlling ER/Golgi Transport

控制 ER/高尔基体运输的蛋白质相互作用

• 批准号: 7924533
• 负责人: JESSE C HAY
• 金额: $ 29.18万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7924533
• 关键词: Address; Adhesions; Back; Biochemical; Biogenesis; Calcium; Calcium ion; Cell physiology; Complex; Confusion; Disease; Docking; Documentation; Drops; Environment; Event; Golgi Apparatus; Imaging Techniques; In Vitro; Individual; Intracellular Membranes; Intrinsic factor; Knowledge; Link; Mammals; Mediating; Membrane; Membrane Fusion; Neuropathy; Organelles; Pathway interactions; Peripheral; Physiological; Play; Process; Protein Family; Protein Secretion; Proteins; Public Health; Pus; Quality Control; Regulation; Role; SNAP receptor; Signal Transduction; Site; Specific qualifier value; Staging; System; Testing; Transmembrane Transport; Transport Vesicles; Tubular formation; Vesicle; Viral; in vivo; interest; particle; prevent; protein complex; protein transport; reconstitution; secretory protein; sensor; trafficking

DESCRIPTION (provided by applicant): Proper cell function and avoidance of disease requires subcellular targeting and fusion of transport vesicles with acceptor membranes. The initial recognition between vesicles and acceptor membranes is mediated by peripheral membrane tether protein complexes, and their specific fusion is mediated by integral membrane SNARE complexes. Although the general function of each of these protein families is established, the protein interactions mediating a tethering event-the specific recognition and adhesion of membranes prior to fusion-are largely unknown and our knowledge of the regulatory relationships linking and integrating tether and SNARE function is very rudimentary. Furthermore, intracellular membrane fusion steps are in many cases regulated by and even dependent upon the release of luminal calcium ions, yet documentation of these important roles in specific transport steps, identification of the machinery mediating calcium release and elucidation of the pathways connecting calcium to tether and SNARE function remain elusive. We developed an in vitro reconstitution system for elucidation of tether and SNARE function in the first membrane fusion step in the secretory pathway, the homotypic fusion of COPII vesicles to generate VTCs, a step that was previously unstudied. We will use our in vitro reconstitution in conjunction with in vivo imaging techniques to address the following specific aims: 1) Test the hypothesis that homotypic COPII vesicle tethering is a multi-layered process involving SNARE-signaled tether recruitment and sequential action multiple tether molecules. 2) Test the hypothesis that calcium negatively regulates homotypic COPII vesicle fusion. 3) Test the hypothesis that the luminal calcium concentration intrinsic to each organelle is an important determinant of membrane transport in the early secretory pathway. RELEVANCE TO PUBLIC HEALTH: The homotypic fusion of COPII vesicles produces VTCs, an organelle with a central role in secretory pathway quality control. Multiple diseases, including severe neuropathies, can result from improper VTC trafficking of misfolded secretory proteins, and VTCs have recently been implicated in the biogenesis of viral particles and other functions. A firm understanding of VTC biogenesis is central to an understanding of the role of VTCs in pathogenic states.

描述（由申请人提供）：适当的细胞功能和疾病的避免需要用受体膜的运输囊泡的亚细胞靶向和融合。囊泡和受体膜之间的初始识别是由周围膜系蛋白蛋白复合物介导的，其特异性融合是由整体膜SNARE复合物介导的。尽管建立了这些蛋白质家族的每个蛋白质的一般功能，但介导束缚事件的蛋白质相互作用 - 融合之前的特定识别和对膜的粘附 - 在很大程度上未知，并且我们对与链球关系的调节关系的了解和积分和SNARE功能相关的调节关系是非常朴素的。此外，在许多情况下，细胞内膜融合步骤受到腔钙离子的释放的调节，但在特定传输步骤中释放了这些重要作用，鉴定钙释放的机械识别以及阐明钙连接到Tether和Snare功能的途径仍然难以捉摸。我们开发了一个体外重构系统，用于在分泌途径的第一个膜融合步骤中阐明系绳和圈圈功能，这是Copii囊泡生成VTC的同型融合，这是以前未研究的一步。我们将结合体内成像技术的体外重构来解决以下特定目的：1）检验以下假设：同型复印囊膜束带是一个多层的过程，涉及涉及的nare签名的绑线募集和顺序作用作用和顺序作用。 2）检验钙对同型复印融合的负调节的假设。 3）检验以下假设：每个细胞器固有的腔钙浓度是早期分泌途径中膜转运的重要决定因素。与公共卫生的相关性：Copii囊泡的同型融合会产生VTC，这是一种在分泌途径质量控制中具有核心作用的细胞器。多种疾病，包括严重的神经病，可能是由于对分泌蛋白错误折叠式分泌蛋白的不当运输可能导致的，而VTC最近与病毒颗粒和其他功能的生物发生有关。对VTC生物发生的坚定理解对于理解VTC在致病状态的作用至关重要。

项目成果

Poliovirus infection transiently increases COPII vesicle budding.

脊髓灰质炎病毒感染暂时增加 COPII 囊泡出芽。

• DOI: 10.1128/jvi.01159-12
• 发表时间: 2012
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Trahey,Meg;Oh,HyungSuk;Cameron,CraigE;Hay,JesseC
• 通讯作者: Hay,JesseC

p115-SNARE interactions: a dynamic cycle of p115 binding monomeric SNARE motifs and releasing assembled bundles.

• DOI: 10.1111/tra.12242
• 发表时间: 2015-02
• 期刊: Traffic (Copenhagen, Denmark)
• 作者: Wang T;Grabski R;Sztul E;Hay JC
• 通讯作者: Hay JC

Transport vesicle uncoating: it's later than you think.

• DOI: 10.3410/b2-47
• 发表时间: 2010
• 期刊: F1000 biology reports
• 作者: Trahey M;Hay JC
• 通讯作者: Hay JC

Reconstitution of COPII vesicle fusion to generate a pre-Golgi intermediate compartment.

• DOI: 10.1083/jcb.200408135
• 发表时间: 2004-12-20
• 期刊: JOURNAL OF CELL BIOLOGY
• 影响因子: 7.8
• 作者: Xu, DL;Hay, JC
• 通讯作者: Hay, JC

mBet3p is required for homotypic COPII vesicle tethering in mammalian cells.

• DOI: 10.1083/jcb.200603044
• 发表时间: 2006-07-31
• 期刊: JOURNAL OF CELL BIOLOGY
• 影响因子: 7.8
• 作者: Yu, Sidney;Satoh, Ayano;Pypaert, Marc;Mullen, Karl;Hay, Jesse C;Ferro-Novick, Susan
• 通讯作者: Ferro-Novick, Susan