Role of beta-arrestins in chemokine receptor signaling

β-抑制蛋白在趋化因子受体信号传导中的作用

• 批准号: 10391496
• 负责人: Adriano Marchese
• 金额: $ 32.05万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-11 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10391496
• 关键词: Adaptor Signaling Protein; Anatomy; Attenuated; Binding; Biochemistry; Biophysics; CXCL12 gene; Cell Culture Techniques; Cell Survival; Cell physiology; Cell surface; Cells; Cellular biology; Cessation of life; Chemotaxis; Complex; Coupling; Data; Disease; Disseminated Malignant Neoplasm; Distant; Focal Adhesion Kinase 1; Focal Adhesions; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Genetic; Health; Heterotrimeric GTP-Binding Proteins; Homing; In Vitro; Integrins; Knowledge; Lead; Ligands; Link; Liver; Lung; MAPK3 gene; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metastatic to; Modeling; Molecular; Molecular Biology; Molecular Conformation; Mutagenesis; Pathway interactions; Phosphorylation; Play; Process; Prognosis; Publishing; RNA Interference; Receptor Signaling; Reporting; Research; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Site; Specific qualifier value; Spectrum Analysis; Stimulus; Surface; Techniques; Testing; Tissues; Vesicle; base; beta-arrestin; biophysical properties; cancer cell; cell motility; chemokine; chemokine receptor; design; innovation; insight; live cell imaging; lymph nodes; novel; overexpression; prevent; protein B; receptor; receptor binding; targeted treatment; therapeutic target; tumor progression

PROJECT SUMMARY The heterotrimeric G protein-coupled receptor (GPCR) C-X-C motif receptor 4 (CXCR4) and its cognate ligand CXCL12 play important roles in health and disease. A large body of evidence indicates that CXCR4 signaling is linked to cancer progression. CXCR4 expression and signaling in cancer correlates with poor prognosis2-5, mainly because cancer cells expressing CXCR4 colonize distant anatomical sites where CXCL12 is located, resulting in metastatic disease, the cause of most cancer related deaths. CXCR4 signaling regulates several aspects of cell physiology linked to cancer progression. This includes directed cell migration and cell survival, which occur via several discrete signaling pathways. Yet the mechanisms remain poorly understood. The focus of this proposal is on the signal transduction mechanisms that regulate CXCR4-mediated chemotaxis towards CXCL12. We recently reported that CXCR4-mediated chemotaxis occurs via a novel mechanism involving a complex formed between endocytic adaptor proteins b-arrestin1 (barr1) and STAM1 (barr1:STAM1). The barr1:STAM1 complex does not act on Akt or ERK-1/2 signaling pathways, but instead is necessary for activating focal adhesion kinase (FAK), which is also necessary for CXCL12 driven chemotaxis. FAK is typically linked to integrin signaling and focal adhesion dynamics, but these aspects of FAK function are not regulated by the barr1:STAM1 complex. Despite our contribution how barr1:STAM1 activates FAK downstream of CXCR4 to promote chemotaxis remains poorly understood. The overall objective of this proposal is to fill in knowledge gaps. Based on our published and preliminary studies we hypothesize that G protein-dependent barr1:STAM1 signaling spatially and temporally controls FAK activity required for CXCR4-dependent chemotaxis. To test this hypothesis we will pursue the following specific aims: Aim 1. To elucidate the role of CXCR4 site-specific phosphorylation on FAK activation; Aim 2. To identify the structural and biophysical properties of the barr1 interaction with STAM1; Aim 3. To elucidate the functional role of the barr1:STAM1 complex in chemotaxis. Because of the mechanistic focus of our proposal we will use cell culture models and other in vitro approaches spanning techniques in cell and molecular biology, genetics, biochemistry and biophysics plus advanced live cell imaging strategies and mass spectrometry approaches. At the conclusion of this project we will have learned novel signal transduction mechanisms by which barr1:STAM1 collaborate to activate FAK to promote chemotaxis. This is significant because it will reveal novel aspects of CXCR4 signaling that could be targeted therapeutically.

项目摘要 异三聚体G蛋白偶联受体（GPCR）C-X-C基序受体4（CXCR4）及其同源配体 CXCL12在健康和疾病中起重要作用。大量证据表明CXCR4信号传导 与癌症进展有关。癌症的CXCR4表达和信号传导与预后不良相关2-5， 主要是因为表达CXCR4的癌细胞在CXCL12所在的遥远的解剖部位定植， 导致转移性疾病，这是大多数与癌症相关的死亡的原因。 CXCR4信号调节几个 与癌症进展有关的细胞生理学方面。这包括定向细胞迁移和细胞存活， 通过几个离散信号通路发生。然而，这些机制仍然知之甚少。重点 该提议的是信号转导机制，该机制调节CXCR4介导的趋化性 CXCL12。我们最近报道说，CXCR4介导的趋化性是通过涉及a的新机制发生的 在内吞衔接蛋白B-arrestin1（Barr1）和Stam1（Barr1：Stam1）之间形成的复合物。这 BARR1：Stam1复合物不对AKT或ERK-1/2信号通路作用，而是必要的 激活局灶性粘附激酶（FAK），这对于CXCL12驱动的趋化性也是必需的。 fak是 通常与整联蛋白信号传导和焦点粘附动力学联系在一起，但是FAK功能的这些方面不是 由Barr1：Stam1复合物调节。尽管我们贡献了Barr1：STAM1如何激活FAK下游 促进趋化性的CXCR4的理解仍然很少。该提议的总体目的是填写 知识差距。基于我们发表的初步研究，我们假设G蛋白依赖性 BARR1：STAM1信号在空间和时间上控制CXCR4依赖性所需的FAK活动 趋化性。为了检验这一假设，我们将追求以下特定目的：目标1。阐明 FAK激活时CXCR4位点特异性磷酸化；目标2。确定结构和生物物理 Barr1与Stam1相互作用的特性；目标3。阐明Barr1的功能作用：stam1 趋化性中的复合物。由于我们的提案的机械重点，我们将使用细胞培养模型，并 其他体外方法涵盖细胞和分子生物学，遗传学，生物化学和 生物物理以及先进的活细胞成像策略和质谱法。结束 这个项目我们将学习Barr1：Stam1合作的新型信号转导机制 激活FAK以促进趋化性。这很重要，因为它将揭示CXCR4的新颖方面 可以针对治疗的信号。