MOLECULAR CONTROL OF CALCIUM INFLUX AT THE ER-PLASMA MEMBRANE JUNCTIONS

ER-血浆膜连接处钙流入的分子控制

• 批准号: 10386835
• 负责人: Yubin Zhou
• 金额: $ 29.18万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386835
• 关键词: Adopted; Animal Model; Biochemical; Biological Process; Biophysics; Calcium; Calcium Channel; Calcium Signaling; Cardiovascular Diseases; Cell membrane; Cells; Cellular Metabolic Process; Clinical; Clinical Medicine; Communication; Complex; Coupling; Data; Development; Disease; Drug Targeting; EF Hand Motifs; Exposure to; Functional disorder; Gene Expression; Generations; Genetic; Goals; Homeostasis; Human; Immunologic Deficiency Syndromes; Kinetics; Knock-out; Knockout Mice; Knowledge; Label; Laboratories; Link; Location; Lymphocyte; Lymphocyte Activation; Mammals; Maps; Membrane; Membrane Fusion; Mission; Modeling; Molecular; Muscle; Muscle Contraction; Neoplasm Metastasis; Pathologic; Pathway interactions; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositol Phosphates; Phosphatidylinositols; Physiological; Physiological Processes; Pilot Projects; Protein Engineering; Protein Family; Proteins; Proteomics; Public Health; Research; Route; STIM1 gene; Severe Combined Immunodeficiency; Shapes; Side; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Site; Stimulus; Structure-Activity Relationship; System; T-Cell Activation; Techniques; Testing; Therapeutic; Therapeutic Intervention; Transgenic Mice; Translating; Tubular Aggregate Myopathies; United States National Institutes of Health; base; clinically relevant; disability; druggable target; extracellular; gain of function mutation; human disease; innovation; insight; mouse model; novel; optogenetics; overexpression; public health relevance; therapeutic target; therapeutically effective; tool; trafficking

Project Summary / Abstract. Store-operated calcium entry (SOCE) constitutes a major calcium entry pathway in mammals to control lymphocyte activation, muscle contraction, gene expression and cell metabolism. The calcium release- activated calcium (CRAC) channel composed of ORAI-STIM represents a prototypical example of SOCE in lymphocytes. The clinical relevance of SOCE is exemplified by two human diseases, the severe combined immunodeficiency (SCID) and tubular aggregate myopathy (TAM), which are caused by loss- or gain-of-function mutations in ORAI1 and STIM1, respectively. Augmented SOCE is also implicated in cardiovascular disorders and cancer metastasis. Therefore, CRAC channel has been pursued as an attractive drug target for therapeutic intervention. Tremendous efforts have been directed to establish ORAI-STIM as the minimal two-component system to couple ER calcium store depletion with calcium influx across the plasma membrane. The regulatory machinery dedicated to the ORAI-STIM signaling, nonetheless, still remains incompletely defined. In this proposal, the PI aims to bridge this critical knowledge gap by unveiling the functions of two novel SOCE modulators, which reside at distinct subcellular locations to act on different steps of ORAI-STIM signaling: the initial activation of STIM within the ER lumen and the later stabilization of ORAI-STIM complexes at ER-PM membrane contact sites (MCS), where the close appositions of two membranes are separated by a gap distance of 10-30 nm. In Aim 1, based on preliminary findings from proteomic profiling of potential STIM1 interactors within the ER lumen, the PI will define how a previously-unrecognized multiple EF-hand protein cooperates with the luminal domain of STIM1 (EFSAM) to shape the activation and deactivation kinetics of SOCE. The PI will employ a new “ER-to-PM” trafficking strategy to expose the luminal domain toward the extracellular side, thereby overcoming a major impediment to studies on the liminal sides of ER-resident signaling proteins. In Aim 2, capitalizing on the discovery of a TMEM family protein as a regulator of calcium influx at ER-PM MCS, the PI will define how this modulator responds to physiological stimuli to remodel the assembly of ER-PM junctions and PIP homeostasis to sustain SOCE. The generation of innovative optogenetic tools and a transgenic mouse model to dissect calcium signaling and protein-PIP interactions will further accelerate our structure-function relationship studies on these novel regulators. Overall, the new mechanistic insights gained through the proposed study will lead to advances in the constantly-revitalized field of calcium signaling, and in parallel, spawn the vibrant field of membrane contact sites. In the long run, discoveries made in the study can be translated into the development of effective therapeutics targeting aberrant calcium and phosphoinositide signaling.

项目摘要/摘要。 库操作钙内流(SOCE)是哺乳动物体内控制钙离子内流的主要途径 淋巴细胞活化、肌肉收缩、基因表达和细胞代谢。钙的释放- 由ORAI-STIM组成的激活钙通道(CRAC)是SOCE的典型代表 淋巴细胞。SOCE的临床相关性以两种人类疾病为例，严重合并 免疫缺陷(SCID)和肾小管聚集性肌病()，由功能丧失或恢复引起 ORAI1和STIM1分别发生突变。增强的SOCE也与心血管疾病有关 和癌症转移。因此，CRAC通道被认为是一个有吸引力的药物治疗靶点 干预。已经做出了巨大努力，将ORAI-STIM建立为最小的两个组成部分 内质网钙库耗竭与跨质膜钙内流的耦合系统。监管机构 然而，专门用于ORAI-STIM信令的机制仍然没有完全定义。 在这项提议中，PI旨在通过揭示两部小说的功能来弥合这一关键的知识鸿沟 SOCE调制器，驻留在不同的亚细胞位置，作用于ORAI-STIM的不同步骤 信号转导：内质网腔内STIM的初始激活和ORAI-STIM复合体的后期稳定 在ER-PM膜接触位置(MCS)，其中两个膜的紧密对位被一个 间隙距离为10-30 nm。在目标1中，基于潜在STIM1的蛋白质组图谱的初步发现 内质网管腔内的相互作用，PI将定义以前未被识别的多EF-手蛋白如何 与STIM1的腔结构域(EFSAM)合作形成激活和失活的动力学 索斯。PI将采用一种新的“ER-to-PM”交易策略，将流明领域暴露给 细胞外侧，从而克服了对内质网常驻患者临界侧研究的主要障碍 信号蛋白。在目标2中，利用TMEM家族蛋白作为钙调节因子的发现 在ER-PM MCS，PI将定义该调制器如何响应生理刺激以重塑 组装内质网-质膜连接和PIP稳态以维持SOCE。创新型光伏发电的产生 分析钙信号和蛋白质-PIP相互作用的工具和转基因小鼠模型将进一步 加快我们对这些新型调节剂的结构-功能关系的研究。 总体而言，通过拟议研究获得的新的机械论见解将导致在 不断振兴的钙信号领域，并同时催生充满活力的膜接触领域 网站。从长远来看，研究中的发现可以转化为有效的开发 针对异常钙和肌醇磷脂信号的治疗。

项目成果

Inside-out Ca(2+) signalling prompted by STIM1 conformational switch.

由 STIM1 构象转换引发的由内而外的 Ca(2 ) 信号传导。

• DOI: 10.1038/ncomms8826
• 发表时间: 2015-07-17
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Ma G;Wei M;He L;Liu C;Wu B;Zhang SL;Jing J;Liang X;Senes A;Tan P;Li S;Sun A;Bi Y;Zhong L;Si H;Shen Y;Li M;Lee MS;Zhou W;Wang J;Wang Y;Zhou Y
• 通讯作者: Zhou Y

Integrated pipeline for inferring the evolutionary history of a gene family embedded in the species tree: a case study on the STIMATE gene family.

用于推断物种树中嵌入的基因家族进化历史的综合管道：STIMATE 基因家族的案例研究。

• DOI: 10.1186/s12859-017-1850-2
• 发表时间: 2017-10-03
• 期刊: BMC bioinformatics
• 影响因子: 3
• 作者: Song J;Zheng S;Nguyen N;Wang Y;Zhou Y;Lin K
• 通讯作者: Lin K

A STIMulating journey into optogenetic engineering.

光学遗传工程的刺激旅程。

• DOI: 10.1016/j.ceca.2020.102197
• 发表时间: 2020-05-04
• 期刊: Cell calcium
• 影响因子: 4
• 作者: Ma G;Zhou Y
• 通讯作者: Zhou Y

TRIM59 promotes breast cancer motility by suppressing p62-selective autophagic degradation of PDCD10.

• DOI: 10.1371/journal.pbio.3000051
• 发表时间: 2018-11
• 期刊: PLoS biology
• 影响因子: 9.8
• 作者: Tan P;Ye Y;He L;Xie J;Jing J;Ma G;Pan H;Han L;Han W;Zhou Y
• 通讯作者: Zhou Y

STIM2 interacts with AMPK and regulates calcium-induced AMPK activation.

• DOI: 10.1096/fj.201801225r
• 发表时间: 2019-03
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Chauhan AS;Liu X;Jing J;Lee H;Yadav RK;Liu J;Zhou Y;Gan B
• 通讯作者: Gan B