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: 10300898
• 负责人: Adriano Marchese
• 金额: $ 1.3万
• 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 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10300898
• 关键词: Adaptor Signaling Protein; Anatomy; Biochemistry; Biophysics; CXCL12 gene; Cell Culture Techniques; Cell Survival; Cell physiology; Cellular biology; Cessation of life; Chemotaxis; Complex; Disease; Disseminated Malignant Neoplasm; Distant; Focal Adhesion Kinase 1; Focal Adhesions; G-Protein-Coupled Receptors; GTP-Binding Proteins; Genetic; Health; Heterotrimeric GTP-Binding Proteins; In Vitro; Integrins; Knowledge; Lead; Ligands; Link; MAPK3 gene; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metastatic to; Modeling; Molecular; Molecular Biology; Phosphorylation; Play; Process; Prognosis; Publishing; Receptor Signaling; Reporting; Role; Signal Pathway; Signal Transduction; Site; Structure; Techniques; Testing; Tissues; base; beta-arrestin; biophysical properties; cancer cell; cell motility; chemokine receptor; live cell imaging; new therapeutic target; novel; overexpression; prevent; protein B; receptor; targeted treatment; 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.

项目总结 异三聚体G蛋白偶联受体C-X-C基序受体4及其同源配体 CXCL12在健康和疾病中发挥着重要作用。大量证据表明，CXCR4信号 与癌症的进展有关。CXCR4在肿瘤中的表达和信号转导与不良预后相关2-5 主要是因为表达CXCR4的癌细胞在CXCL12所在的远处解剖部位定植， 导致转移性疾病，这是大多数癌症相关死亡的原因。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， 这通过几个离散的信号通路发生。然而，人们对这些机制仍然知之甚少。焦点 这一建议的重点是调节CXCR4介导的趋化作用的信号转导机制 CXCL12。我们最近报道，CXCR4介导的趋化作用是通过一种新的机制发生的，涉及一种 内吞接头蛋白b-arrestin1(Barr1)与STAM1(barr1：STAM1)形成复合体。这个 Barr1：STAM1复合体不作用于Akt或ERK-1/2信号通路，而是必需的 激活粘着斑激酶(FAK)，这也是CXCL12驱动趋化所必需的。FAK是 通常与整合素信号和焦点黏附动力学有关，但FAK功能的这些方面不是 受barr1：STAM1复合体调控。尽管我们做出了贡献，但barr1：STAM1如何激活FAK CXCR4下游促进趋化作用的机制仍知之甚少。这样做的总体目标是 建议是填补知识空白。根据我们已发表的和初步的研究，我们假设G 蛋白质依赖的Barr1：STAM1信号在空间和时间上控制CXCR4-所需的FAK活性 依赖性趋化性。为了验证这一假设，我们将追求以下具体目标：目标1.阐明 CXCR4位点特异性磷酸化在FAK激活中的作用；目的2.鉴定结构和 Barr1与STAM1相互作用的生物物理性质；目的3.阐明Barr1与STAM1的功能作用 Barr1：趋化作用中的STAM1复合体。由于我们提案的机械性重点，我们将使用细胞培养 模型和其他体外方法跨越细胞和分子生物学、遗传学、 生物化学和生物物理学加上先进的活细胞成像策略和质谱学方法。在… 这个项目的结论是，我们将学习到新的信号转导机制，通过这些机制 Barr1：STAM1协同激活FAK促进趋化作用。这一点意义重大，因为它将揭示小说 CXCR4信号的某些方面可以作为治疗靶点。