Mechanisms and function of endosome-derived tubular carriers

内体衍生的管状载体的机制和功能

• 批准号: 10000963
• 负责人: Steven H Caplan
• 金额: $ 31.84万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10000963
• 关键词: Address; Atherosclerosis; Biogenesis; Biological; Biological Assay; Biophysics; Cell membrane; Cell physiology; Cells; Complex; Data; Diabetes Mellitus; Disease; Elements; Endosomes; Generations; Golgi Apparatus; Grant; Health; Homeostasis; Knowledge; Laboratories; Malignant Neoplasms; Membrane; Membrane Biology; Microscopy; Molecular; Normal Cell; Organelles; Pathway interactions; Phosphatidic Acid; Physiological; Play; Process; Proteins; Receptor Signaling; Recycling; Regulation; Research; Resolution; Role; Sorting - Cell Movement; Structure; System; Techniques; Tertiary Protein Structure; Tubular formation; base; neglect; novel; receptor; receptor recycling; recruit; sorting nexins; spatiotemporal; structural biology; trafficking; trans-Golgi Network

Endocytic trafficking is central to normal cell function, and dysregulation is the underlying cause for diseases as diverse as atherosclerosis, diabetes and cancer. My laboratory has contributed to understanding the mechanisms by which membranes and receptors are recycled via the endocytic recycling compartment (ERC) to the plasma membrane (PM). Endocytic recycling remains one of the least studied endocytic pathways; in particular, the involvement of membrane tubules in the recycling process is poorly understood, and the roles of the various forms of tubular endosomes, and the mechanisms by which these structures are generated and undergo vesiculation remain a major unanswered question. There are multiple `types' of endosome-derived tubular carriers (EDTC), including sorting nexin-BAR (SNX-BAR) domain and retromer-derived tubules, and networks of endosomal tubules, such as tubular recycling endosomes (TRE) decorated by MICAL-L1, Syndapin2 (Synd2) and Eps15 Homology Domain (EHD) proteins. Little is known about how these apparently different EDTC integrate their functions, or even whether retromer and SXN-BAR EDTC are distinct from TRE. Recent studies demonstrate that EHD proteins and MICAL-L1 interact with components of the retromer complex, suggesting that these tubular networks are related. Our central hypothesis is that overlapping and distinct tubular membranes coordinate transport from endosomes to the PM and the Golgi. In this proposal, we will uncover the functional and physical relationships between TRE, retromer and SNX-BAR-derived EDTC. Moreover, we will focus on a mechanistic understanding of the mode by which EDTC are generated, and how they undergo vesiculation to promote transport within the cell. Aim 1: To examine the relationship and functional roles of EDTC. Tubular endosomes play major roles in endocytic membrane trafficking. EDTC include a number of retromer-containing structures, retromer-independent tubules generated by SNX-BAR domain proteins, and TRE decorated by MICAL-L1, the BAR domain protein Synd2, and EHD proteins. We will examine the cross-talk and cross-regulation of these pathways with regard to the generation and fission of recycling tubules, and the control of endocytic recycling. Aim 2: To define and elucidate the spatio-temporal regulation and mechanism of TRE biogenesis and vesiculation. Our working hypothesis is that phosphatidic acid generation leads to recruitment of the membrane hub, MICAL-L1 and the F-BAR protein Synd2, to generate and remodel TRE. We further hypothesize that EHD3 plays an essential role in this process by stabilizing MICAL-L1-Synd2 interactions. Our studies on the cellular, organellar, molecular and atomic levels will use techniques ranging from structural biology to super-resolution microscopy and novel biophysical vesiculation assays, providing crucial knowledge of the functional role of the poorly understood TRE and retromer-SNX-BAR derived tubules, and their biogenesis and vesiculation.

内吞运输是正常细胞功能的核心，失调是导致的根本原因 像动脉粥样硬化，糖尿病和癌症一样多样化的疾病。我的实验室为 了解通过内吞膜和受体回收膜和受体的机制 回收室（ERC）到质膜（PM）。内吞回收仍然是最少的 研究了内吞途径；特别是，膜小管在回收过程中的参与是 理解不佳，以及各种形式的管状内体的作用，以及所在的机制 这些结构是生成的，并且经过囊泡仍然是一个主要的未解决问题。有 内体衍生的管状载体（EDTC）多种“类型”，包括对Nexin-Bar（SNX-BAR）进行排序 域和逆转录小管，以及内体小管的网络，例如管状回收 由Mical-L1，Syndapin2（Synd2）和EPS15同源域（EHD）装饰的内体（TRE）（TRE） 蛋白质。对于这些显然不同的EDTC如何整合其功能，甚至几乎没有了解 逆转录和SXN-BAR EDTC是否与TRE不同。最近的研究表明EHD 蛋白质和mical-L1与逆转录络合物的成分相互作用，表明这些管状 网络是相关的。我们的中心假设是重叠和独特的管状膜坐标 从内体到PM和高尔基体的运输。在此提案中，我们将发现功能和 TRE，缩回和SNX衍生的EDTC之间的物理关系。此外，我们将重点放在 对EDTC产生的模式的机械理解，以及它们如何进行囊泡 促进电池内的运输。目标1：检查EDTC的关系和功能作用。 管状内体在内吞膜贩运中起主要作用。 EDTC包括许多 含回逆转录的结构，由SNX-BAR结构蛋白产生的逆转录者无关的小管， 由Mical-L1，bar域蛋白Synd2和EHD蛋白装饰的TRE。我们将检查 这些途径在回收的产生和裂变方面的盘问和交叉调节 小管和内吞回收的控制。目标2：定义和阐明时空 TRE生物发生和囊泡的调节和机制。我们工作的假设是 磷脂酸的产生可导致膜枢纽，mical-L1和F-BAR蛋白的募集 Synd2，生成和重塑TRE。我们进一步假设EHD3在此中起着至关重要的作用 通过稳定Mical-L1-SynD2相互作用来过程。我们对细胞，细胞器，分子和 原子水平将使用从结构生物学到超分辨率显微镜和新颖的技术 生物物理囊泡测定法，提供对知之甚少的功能作用的重要知识 TRE和反术-SNX-BAR衍生的小管及其生物发生和囊泡。

项目成果

Differential requirements for the Eps15 homology domain proteins EHD4 and EHD2 in the regulation of mammalian ciliogenesis.

• DOI: 10.1111/tra.12845
• 发表时间: 2022-07
• 期刊: TRAFFIC
• 影响因子: 4.5
• 作者: Jones, Tyler;Naslavsky, Naava;Caplan, Steve
• 通讯作者: Caplan, Steve

The retromer complex regulates C. elegans development and mammalian ciliogenesis.

逆转录酶复合体调节线虫发育和哺乳动物纤毛发生。

• DOI: 10.1242/jcs.259396
• 发表时间: 2022
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Xie,Shuwei;Dierlam,Carter;Smith,Ellie;Duran,Ramon;Williams,Allana;Davis,Angelina;Mathew,Danita;Naslavsky,Naava;Iyer,Jyoti;Caplan,Steve
• 通讯作者: Caplan,Steve

Tying trafficking to fusion and fission at the mighty mitochondria.

• DOI: 10.1111/tra.12573
• 发表时间: 2018-08
• 期刊: Traffic (Copenhagen, Denmark)
• 作者: Farmer T;Naslavsky N;Caplan S
• 通讯作者: Caplan S

MICAL2PV suppresses the formation of tunneling nanotubes and modulates mitochondrial trafficking.

• DOI: 10.15252/embr.202052006
• 发表时间: 2021-07-05
• 期刊: EMBO reports
• 影响因子: 7.7
• 作者: Wang F;Chen X;Cheng H;Song L;Liu J;Caplan S;Zhu L;Wu JY
• 通讯作者: Wu JY

Coronin2A links actin-based endosomal processes to the EHD1 fission machinery.

• DOI: 10.1091/mbc.e21-12-0624
• 发表时间: 2022-10-01
• 期刊: MOLECULAR BIOLOGY OF THE CELL
• 影响因子: 3.3
• 作者: Dhawan, Kanika;Naslavsky, Naava;Caplan, Steve
• 通讯作者: Caplan, Steve