The function of MEKK3 interaction with CCM2

MEKK3与CCM2相互作用的功能

基本信息

批准号：
8863345
负责人：
Titus Jonathon Boggon
金额：
$ 32.05万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8863345
关键词：
Address Affect Binding Binding Sites Biochemical Biological Assay Blood Vessels Brain CCM1 gene Cavernous Malformation Cells Cerebrum Chronic Headaches Complex Data Defect Dimerization Disease Endothelial Cells Epilepsy Figs - dietary Functional disorder Gene Targeting Genes Genetic Goals Laboratories Lesion Link MAP Kinase Kinase Kinase MEKKs Maps Mediating Mitogen-Activated Protein Kinase Kinases Mitogen-Activated Protein Kinases Molecular Molecular Genetics Mutation Neurologic Pathology Pathway interactions Patients Phenotype Phosphorylation Phosphotransferases Physiological Physiology Play Population Process Protein Kinase Proteins Recruitment Activity Regulation Research Research Design Role Seizures Signal Transduction Stroke Structure Subgroup Tertiary Protein Structure Testing Vascular System base biochemical tools biophysical tools improved in vitro activity in vivo loss of function mouse model protein complex public health relevance vascular abnormality

项目摘要

DESCRIPTION (provided by applicant): Cerebral Cavernous Malformations (CCMs) are brain vascular lesions estimated to affect up to 0.5% of the population. CCM patients can suffer chronic headaches, epilepsy, seizures, stroke and focal neurological deficits. Three disease-associated genes, which encode CCM1/KRIT1, CCM2/malcavernin/OSM, and CCM3/PDCD10, respectively, have been identified. These CCM proteins are known to form a multi-protein complex (the CCM complex signaling platform) and a loss of function of any one of these proteins leads to CCM pathology. Despite major progress in our understanding of the genetics and molecular functions of CCM proteins in mouse models, precisely how CCM is developed in patients with mutations in CCM genes remains largely unclear. One of the known binding partners of the CCM complex is a mitogen-activated protein kinase kinase kinase, MEKK3. Importantly, the role of MEKK3 in the vascular system appears to be overlapping with that of CCM proteins, especially that of CCM2. Therefore the goal of this project is to understand the structural requirement and functional consequences of MEKK3:CCM2 interaction and the associated changes in downstream signaling that may impact the critical vasculature phenotypes associated with CCM disease. Towards this end, our studies are designed to comprehensively address our central hypothesis that recruitment of MEKK3 by CCM2 is critical for vascular integrity. We will address this hypothesis in three Aims. In Aim 1 we will define the structural and functional mechanisms for MEKK3 recruitment to CCM2. This aim will use structural, biochemical and biophysical tools to provide the basic molecular level framework for how MEKK3 is recruited to the CCM complex by its interaction with CCM2. In Aim 2 we will investigate the role of CCM2 in regulation of MEKK3 signaling by probing MEKK3 regulation by CCM2 using biochemical and cell based assays. We will also investigate the specific phosphorylation targets of the MEKK3:CCM2 complex and investigate the target genes of the complex. In Aim 3 we will discover the in vivo functional role of MEKK3 recruitment to CCM2. We utilize structure-directed in vivo studies, in which we specifically disrupt the MEKK3:CCM2 interaction without compromising the overall activity of MEKK3, to test whether critical vasculature phenotypes associated with CCM disease, and associated changes in downstream signaling, result from loss of the MEKK3:CCM2 interaction. Overall, we expect that the studies we propose will define the functional importance of MEKK3 interaction with CCM2 and resolve whether the vascular pathology associated with CCM disease result from loss of the MEKK3:CCM2 interaction and associated changes in downstream signaling.

描述（由申请人提供）：脑海绵状血管畸形 (CCM) 是一种脑血管病变，估计影响高达 0.5% 的 CCM 患者，可能患有慢性头痛、癫痫、癫痫发作、中风和局灶性神经功能缺损。，分别编码 CCM1/KRIT1、CCM2/malcavernin/OSM 和 CCM3/PDCD10，这些 CCM 已被识别。尽管我们对小鼠 CCM 蛋白的遗传学和分子功能的理解取得了重大进展，但已知蛋白质会形成多蛋白复合物（CCM 复合物信号平台），并且其中任何一种蛋白质的功能丧失都会导致 CCM 病理学。在 CCM 基因突变的患者中，CCM 是如何形成的，目前仍不清楚。CCM 复合物的已知结合伴侣之一是丝裂原激活的蛋白激酶激酶 MEKK3。重要的是，MEKK3 在血管系统中的作用。看来与 CCM 蛋白的重叠，尤其是 CCM2 的重叠，因此该项目的目标是了解 MEKK3:CCM2 相互作用的结构要求和功能后果，以及可能影响与 CCM 相关的关键脉管系统表型的下游信号传导的相关变化。为此，我们的研究旨在全面解决我们的中心假设，即 CCM2 招募 MEKK3 对于血管完整性至关重要，我们将在目标 1 中定义这一假设。 MEKK3 招募到 CCM2 的结构和功能机制该目标将使用结构、生物化学和生物物理工具来提供 MEKK3 如何通过与 CCM2 相互作用招募到 CCM 复合物的基本分子水平框架。通过使用生化和基于细胞的检测来探究 CCM2 对 MEKK3 的调节，我们还将研究 MEKK3:CCM2 复合物的特定磷酸化靶点并研究 CCM2 对 MEKK3 信号传导的调节。在目标 3 中，我们将发现 MEKK3 募集到 CCM2 的体内功能作用，我们利用结构导向的体内研究，在不影响 MEKK3 整体活性的情况下特异性破坏 MEKK3:CCM2 相互作用。测试与 CCM 疾病相关的关键脉管系统表型以及下游信号传导的相关变化是否是由于 MEKK3:CCM2 相互作用的丧失所致。总体而言，我们期望我们提出的研究将定义 MEKK3:CCM2 相互作用的功能重要性。 MEKK3 与 CCM2 的相互作用，并解决与 CCM 疾病相关的血管病理学是否是由 MEKK3:CCM2 相互作用的丧失以及下游信号传导的相关变化引起的。