Cell Signaling by Protease-activated G Protein-coupled Receptors

蛋白酶激活的 G 蛋白偶联受体的细胞信号传导

• 批准号: 10371096
• 负责人: Joann Trejo
• 金额: $ 46.5万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371096
• 关键词: Arrestins; Binding; Biology; Bypass; Complex; Disease; Drug Targeting; Endosomes; Endothelium; Family member; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Goals; Inflammation; Knowledge; Laboratories; Malignant Neoplasms; Mediating; Mediator of activation protein; Molecular; National Institute of General Medical Sciences; PAR-1 Receptor; Pathway interactions; Peptide Hydrolases; Permeability; Phosphorylation; Physiology; Proteins; Regulation; Research; Research Support; Role; Signal Transduction; Sorting - Cell Movement; Therapeutic; Ubiquitin; Ubiquitination; Work; drug development; experience; grasp; malignant breast neoplasm; new therapeutic target; novel; p38 Mitogen Activated Protein Kinase; programs; purinoceptor P2Y1; receptor; receptor function; trafficking; tumor progression

Summary The overall goals of our NIGMS-supported research program have been to delineate the regulatory mechanisms that control signaling by protease-activated receptor-1 (PAR1) and closely-related family members in normal physiology and disease. Our efforts have led to the discovery that ubiquitination of PAR1 promotes p38 mitogen-activated protein kinase (MAPK) signaling from endosomes and not lysosomal degradation. Ubiquitination of the P2Y1 receptor functions similarly. This work further revealed a distinct function for ubiquitination of PAR1 in p38-mediated endothelial barrier disruption. We then discovered a novel lysosomal pathway for PAR1 that is also relevant to other G protein-coupled receptors (GPCRs). Contrary to conventional view, we found that ubiquitination of certain GPCRs and canonical ubiquitin-binding ESCRTs are not required for receptor lysosomal degradation. We then identified ALG- interacting protein (ALIX) and arrestin-related domain containing protein-3 (ARRDC3) as key mediators of GPCR lysosomal sorting. The ALIX-dependent pathway bypasses the requirement for GPCR ubiquitination and is distinct from the canonical endosomal sorting complexes required for transport (ESCRT) lysosomal pathway. The ALIX and ARRDC3 pathway also appears to be dysregulated in cancer. The central premise for the proposed studies is that ubiquitination offers novel and diverse mechanisms for regulation of GPCR biology. A thorough understanding of the mechanism by which key regulators and mediators of ubiquitination regulate GPCR signaling and trafficking is essential for understanding dysregulated mechanisms in disease and for identifying new drug targets. The proposed studies will allow my research group to grasp a thorough understanding of novel concepts that we pioneered - that ubiquitination of GPCRs regulates signaling in normal physiology and is disrupted in disease. Our studies have raised many exciting questions and the most transformative research will be pursued. The p38 MAPK mediates inflammation, however it is not known how GPCR-activated, ubiquitin-driven p38 signaling promotes endothelial barrier permeability, a key feature of inflammation, and will be determined. There is a limited understanding of how GPCR endosomal signaling is regulated and is a challenge. Thus, we will define the mechanisms that regulate GPCR-activated, ubiquitin-driven p38 endosomal signaling. Several GPCRs sort through the ALIX-dependent pathway rather than canonical ESCRT, however, the key steps that segregate GPCRs into distinct lysosomal pathways are not known and will be delineated. ARRCD3 expression is either lost or suppressed in breast cancer, however it is not clear how ARRDC3 impacts PAR1 function or contributes to cancer progression and will be examined. We are well equipped to perform these studies given my laboratory's expertise and experience delineating the function of ubiquitination in regulation of GPCR signaling and trafficking in normal physiology and disease.

概括 我们诺格姆斯支持的研究计划的总体目标是描述监管 通过蛋白酶激活的受体1（PAR1）和密切相关家族来控制信号的机制 正常生理和疾病的成员。我们的努力导致发现了 PAR1促进p38有丝分裂原激活的蛋白激酶（MAPK）来自内体的信号传导，而不是 溶酶体降解。 P2Y1受体的泛素化功能类似。这项工作进一步揭示了 p38介导的内皮屏障破坏中PAR1泛素化的独特功能。然后我们 发现了一种新型PAR1的溶酶体途径，它也与其他G蛋白偶联受体有关 （GPCR）。与传统观点相反，我们发现某些GPCR和规范的泛素化 受体溶酶体降解不需要泛素结合的大肠杆菌。然后，我们确定了ALG- 相互作用的蛋白（ALIX）和含有蛋白质3（ARRDC3）的与逮捕蛋白相关结构域作为关键介体 GPCR溶酶体分选。 ALIX依赖性途径绕过GPCR的要求 泛素化，与运输所需的规范内体分类复合物不同 （ESCRT）溶酶体途径。 ALIX和ARRDC3途径在癌症中似乎也失调。 拟议研究的中心前提是泛素化提供了新颖而多样的机制 用于调节GPCR生物学。对关键监管机构和 泛素化介体调节GPCR信号传导和贩运​​对于理解至关重要 疾病和鉴定新药物靶标的机制失调。拟议的研究将允许 我的研究小组掌握了对我们开创的新颖概念的彻底理解 - GPCR的泛素化调节正常生理中的信号传导，并在疾病中受到干扰。我们的研究 已经提出了许多令人兴奋的问题，将进行最具变革性的研究。 p38 MAPK介导炎症，但是尚不清楚GPCR激活泛素驱动的p38 信号传导促进内皮屏障渗透率，这是炎症的关键特征，并将确定。 人们对GPCR内体信号如何受到调节的方式有限，这是一个挑战。因此， 我们将定义调节GPCR激活，泛素驱动的p38内体信号传导的机制。 但是，几个GPCR通过ALIX依赖性途径进行排序，而不是规范的ESCRT，但是 将GPCR分离为不同的溶酶体途径的关键步骤尚不清楚，并且将被描述。 ARRCD3表达在乳腺癌中丢失或抑制，但是尚不清楚ARRDC3如何 影响PAR1功能或有助于癌症进展，并将检查。我们设备齐全 鉴于我的实验室的专业知识和经验来划定这些研究的作用 正常生理和疾病中GPCR信号传导和贩运​​调节中的泛素化。