Signal Integration from Membranes to the Actin Cytoskeleton

从膜到肌动蛋白细胞骨架的信号整合

基本信息

批准号：
10623679
负责人：
Baoyu Chen
金额：
$ 40.65万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10623679
关键词：
Actins Address Binding Binding Sites Biochemistry Biology Cell membrane Collaborations Complex Cryoelectron Microscopy Cytoskeleton DNA Sequence Alteration Defect Development Disease Eukaryotic Cell Fluorescence Fluorescence Microscopy Foundations Guanosine Triphosphate Phosphohydrolases Human Pathology Immune Immunologic Deficiency Syndromes Individual Infection Knowledge Laboratories Ligands Link Malignant Neoplasms Mass Spectrum Analysis Mediating Membrane Membrane Proteins Monomeric GTP-Binding Proteins Morphogenesis Mutation Names Neoplasm Metastasis Neurodevelopmental Disorder Neurons Oxidation-Reduction Parasitic infection Patients Phosphatidylinositols Play Polymers Process Protein Engineering Protein Family Proteins Regulation Research Role Series Signal Transduction Syndrome Therapeutic Vesicle WAVE protein Wiskott-Aldrich Syndrome Work axon guidance cell motility experimental study field study human disease immune activation interest novel particle pathogen polymerization receptor single molecule trafficking vasodilator-stimulated phosphoprotein

项目摘要

My laboratory has a long-term interest in understanding the basic mechanisms underlying actin cytoskeleton regulation and how these mechanisms drive fundamental processes such as cell migration, vesicle trafficking, immune cell activation, and neuron morphogenesis. We are particularly interested in solving the regulation mechanisms of the Wiskott-Aldrich Syndrome Protein (WASP) family proteins. These proteins play a central role in linking membrane signals to the Arp2/3-mediated actin polymerization throughout eukaryotic cells. Ge- netic mutation or misregulation of these proteins is heavily involved in human diseases, including various neu- rodevelopmental disorders, immune syndromes, pathogen infections, and cancer. Despite their importance, the regulation mechanisms of most WASP-family proteins are poorly understood, posing a large barrier to un- derstanding their roles in biology and human pathology and identifying new avenues of treating related dis- eases. Our recent studies have made a series of pivotal contributions to the understanding of the WASP-family protein WAVE and its role in disease. WAVE is incorporated in a large protein assembly named the WAVE Regulatory Complex (WRC), which is essential to polymerizing actin at the plasma membrane. We have deter- mined how WRC is activated by two distinct small GTPases, Rac1 and Arf, how disease-related mutations dis- rupt WRC activation, and how WRC interacts with various ligands, including the neuronal receptor HPO-30, the redox regulator p47phox, and the actin regulator Ena/VASP/UNC-34. In the next five years, we will continue to move the field vertically by addressing a series of new, important questions about WAVE regulation. In parallel, we will start exploring the mechanisms of two other WASP-family proteins, WHAMM and JMY, for which very little is known mechanistically despite their essential roles in regulating actin assembly at endomembranes. We will combine protein engineering, quantitative biochemistry and mass spectrometry, single molecule fluores- cence microscopy, and single particle cryogenic electron microscopy (cryo-EM) to determine how WRC is acti- vated by Arf binding, how distinct Arf and Rac1 binding sites establish cooperativity, and how WRC interacts with several novel ligands important to neuron morphogenesis, axon guidance, and parasite infection. In addi- tion to dissecting mechanisms of individual ligands, we will establish membrane-based single-molecule fluores- cence experiments to determine the mechanisms by which multiple ligands, including inositol phospholipids, GTPases, and membrane proteins, cooperatively activate the WRC in a context closely resembling cell mem- branes. Furthermore, we will determine how WHAMM and JMY are regulated by binding to various ligands. Our work will provide a comprehensive mechanistic framework for understanding WASP family protein regula- tion. This knowledge will be broadly useful to different fields studying actin-related processes. In collaboration with various biologists and clinicians, our work will reveal how mutations in WASP-family proteins derived from patients disrupt function and provide new ideas and targets for the development of novel intervening agents.

我的实验室对理解肌动蛋白细胞骨架的基本机制具有长期兴趣调节以及这些机制如何驱动基本过程，例如细胞迁移，囊泡运输，免疫细胞激活和神经元形态发生。我们对解决法规特别感兴趣 Wiskott-Aldrich综合征蛋白（WASP）家族蛋白的机制。这些蛋白质发挥了中心在整个真核细胞中将膜信号与ARP2/3介导的肌动蛋白聚合联系起来的作用。 ge- 这些蛋白质的网络突变或不调节与人类疾病有关，包括各种神经疾病竞技疾病，免疫综合征，病原体感染和癌症。尽管它们的重要性大多数黄蜂家庭蛋白的调节机制知之甚少，构成了一个巨大的障碍阐明它们在生物学和人类病理学中的作用，并确定治疗相关疾病的新途径轻松。我们最近的研究为理解黄蜂家庭做出了一系列关键贡献蛋白质波及其在疾病中的作用。波浪被纳入名为波的大蛋白质组件中调节复合物（WRC），这对于在质膜上聚合肌动蛋白至关重要。我们有阻止 - 开采了两个不同的小gtpases Rac1和ARF激活WRC的方式，与疾病相关的突变如何分散破裂的WRC激活以及WRC如何与各种配体相互作用，包括神经元受体HPO-30，氧化还原调节剂P47Phox和肌动蛋白调节剂ENA/VASP/UNC-34。在接下来的五年中，我们将继续通过解决有关波浪调节的一系列新的重要问题，垂直移动字段。并联，我们将开始探索另外两个黄蜂家族蛋白Whamm和JMY的机制，为此非常尽管在调节内膜的肌动蛋白组件中起着至关重要的作用，但在机械上几乎没有知名度。我们将结合蛋白质工程，定量生物化学和质谱法，单分子荧光 - Cence显微镜和单个颗粒低温电子显微镜（Cryo-EM），以确定WRC的作用通过ARF结合，不同的ARF和Rac1结合位点如何建立合作性以及WRC如何相互作用有几种对神经元形态发生，轴突引导和寄生虫感染至关重要的新型配体。加上为了解剖单个配体的机制，我们将建立基于膜的单分子荧光 - CENCE实验确定多种配体（包括肌醇磷脂）的机制 GTPases和膜蛋白在上下文中协同激活WRC，非常相似麸皮。此外，我们将确定如何通过与各种配体结合来调节WHAMM和JMY。我们的工作将为理解黄蜂家族蛋白质调节提供一个全面的机械框架。 tion。这些知识将对研究与肌动蛋白相关的过程的不同领域广泛有用。合作对于各种生物学家和临床医生，我们的工作将揭示黄蜂与蛋白质中的突变如何从患者破坏功能，并为开发新型中间药物提供新的想法和目标。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genetics of mirror movements identifies a multifunctional complex required for Netrin-1 guidance and lateralization of motor control.

DOI：
10.1126/sciadv.add5501
发表时间：
2023-05-12
期刊：
Science advances
影响因子：
13.6
作者：
通讯作者：

A two-step actin polymerization mechanism drives dendrite branching.

DOI：
10.1186/s13064-021-00154-0
发表时间：
2021-07-19
期刊：
Neural development
影响因子：
3.6
作者：
Shi R;Kramer DA;Chen B;Shen K
通讯作者：
Shen K

WAVE regulatory complex.

DOI：
10.1016/j.cub.2021.01.086
发表时间：
2021-05-24
期刊：
Current biology : CB
影响因子：
0
作者：
Rottner K;Stradal TEB;Chen B
通讯作者：
Chen B

Structural basis for Retriever-SNX17 assembly and endosomal sorting.

Retriever-SNX17 组装和内体分选的结构基础。

DOI：
10.1101/2024.03.12.584676
发表时间：
2024
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Singla,Amika;Boesch,DanielJ;JoyceFung,HoYee;Ngoka,Chigozie;Enriquez,AveryS;Song,Ran;Kramer,DanielA;Han,Yan;Juneja,Puneet;Billadeau,DanielD;Bai,Xiaochen;Chen,Zhe;Turer,EmreE;Burstein,Ezra;Chen,Baoyu
通讯作者：
Chen,Baoyu

PPP2R1A regulates migration persistence through the NHSL1-containing WAVE Shell Complex.

DOI：
10.1038/s41467-023-39276-w
发表时间：
2023-06-15
期刊：
NATURE COMMUNICATIONS
影响因子：
16.6
作者：
Wang, Yanan;Chiappetta, Giovanni;Guerois, Raphael;Liu, Yijun;Romero, Stephane;Boesch, Daniel J.;Krause, Matthias;Dessalles, Claire A.;Babataheri, Avin;Barakat, Abdul I.;Chen, Baoyu;Vinh, Joelle;Polesskaya, Anna;Gautreau, Alexis M.
通讯作者：
Gautreau, Alexis M.