Transcription regulation & interaction of ORL1 and GAIP

转录调控

• 批准号: 6728956
• 负责人: Pamela P. Palmer
• 金额: $ 24.52万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6728956
• 关键词: DNA footprinting; RNA interference; biological signal transduction; cell surface receptors; gel mobility shift assay; gene expression; gene expression profiling; genetic promoter element; genetic regulation; genetic transcription; nociceptin; northern blottings; opioid receptor; polymerase chain reaction; posttranscriptional RNA processing; receptor expression; site directed mutagenesis; tissue /cell culture; transfection /expression vector

DESCRIPTION (provided by applicant): The opioid-receptor-like (ORL1) receptor, like the classical mu, delta, and kappa opioid receptors, belongs to the G protein-coupled, seven-transmembrane receptor superfamily. Binding of its endogenous peptide ligand nociceptin/orphanin FQ (NOC/OFQ) to ORL1 initiates a G protein-mediated signal transduction pathway, which fulfills essential functions in regulation of nociception, long-term potentiation and response to stress. ORL1 is expressed in multiple alternative splicing forms in a tissue-, cell-type-, and development-specific manner. However, the molecular mechanisms underlying the regulation of the spatially and temporally differential expression of ORL1 are poorly understood and have not been thoroughly investigated. Furthermore, regulators of G protein signaling (RGS) are now believed to play a crucial role in opioid recetor/G protein-mediated signaling. The role of RGS proteins in ORL1 signal transduction is a main focus of this grant. The discovery by this laboratory that the human ORL1 and GAIP (G alpha interacting protein, also named RGS19) genes are linked and their transcriptions are controlled by a shared bi-directional promoter, as well as the new finding that an activation of ORL1 by NOC/OFQ up-regulates GAIP transcription and that GAIP negatively regulates ORL1 signal transduction, have suggested a significant functional role of GAIP in regulating ORL1 gene expression and function. The tight linkage of ORL1 and GAIP genes provides a unique basis to study the transcription regulation of ORL1 and GAIP, and to study the functional role of GAIP and other RGSs in ORL1 signaling. This grant application proposes a hypothesis that GAIP not only physically couples to the ORL1 gene, but also plays a crucial role in regulating ORL1 gene expression and signal transduction. To examine the hypothesis and establish the functional role of GAIP in regulating ORL1, this application proposes specific aims to investigate how ORL1 specifically regulates GAIP transcription and activity, and in turn, how GAIP can specifically modulate ORL1 expression and function. This application also proposes to examine the selectivity of GAIP in regulating ORL1 signaling by comparing with other RGS proteins. Finally, this study also aims to further analyze the functional activities of the key promoter regions of ORL1 and GAIP genes, hence further exploring the mechanisms that control the coordinated but differential cell-type-specific expression of ORL1 and GAIP genes and their alternative splicing variants. This proposed research will help us to understand the regulatory role of GAIP and RGSs in ORL1 signal transduction and gene expression, lay the basis for a novel strategy of modulating NOC/OFQ and ORL1 functions by manipulating GAIP (RGS) gene expression and activity, and therefore, may provide strategic targets for therapeutic applications in pain management and other neurological and mental disorders related to the ORL1 system.

描述（由申请人提供）：阿片类药物样（ORL1）受体，例如经典的MU，Delta和Kappa阿片受体，属于G蛋白偶联，七跨膜受体超级家族。其内源性肽配体Nocptin/Orphanin FQ（NOC/OFQ）与ORL1的结合启动G蛋白介导的信号转导途径，从而实现了调节伤害感受，长期增强应力和应力反应的基本功能。 ORL1以多种替代剪接形式表达，以组织，细胞类型和开发特异性方式表达。但是，对ORL1的空间和时间差异表达调节的分子机制知之甚少，尚未彻底研究。此外，现在据信G蛋白信号传导（RGS）的调节剂在阿片类药物的蛋白质/G蛋白介导的信号传导中起着至关重要的作用。 RGS蛋白在ORL1信号转导中的作用是该赠款的主要重点。 The discovery by this laboratory that the human ORL1 and GAIP (G alpha interacting protein, also named RGS19) genes are linked and their transcriptions are controlled by a shared bi-directional promoter, as well as the new finding that an activation of ORL1 by NOC/OFQ up-regulates GAIP transcription and that GAIP negatively regulates ORL1 signal transduction, have suggested a significant functional role of GAIP in调节ORL1基因表达和功能。 ORL1和GAIP基因的紧密连接为研究ORL1和GAIP的转录调控提供了独特的基础，并研究了GAIP和其他RGSS在ORL1信号传导中的功能作用。该赠款的应用提出了一个假设，即GAIP不仅在物理上与ORL1基因伴侣，而且在调节ORL1基因表达和信号转导中起着至关重要的作用。为了检查假设并确定GAIP在调节ORL1中的功能作用，该应用程序提出了特定的目的，旨在研究O​​RL1如何专门调节GAIP转录和活动，进而调节GAIP如何专门调节ORL1的表达和功能。该应用程序还建议通过与其他RGS蛋白进行比较来检查GAIP在调节ORL1信号传导中的选择性。最后，这项研究还旨在进一步分析ORL1和GAIP基因的关键启动子区域的功能活性，从而进一步探索控制ORL1和GAIP基因及其替代旋转变体的配位但差异细胞特异性表达的机制。这项拟议的研究将帮助我们了解GAIP和RGSS在ORL1信号转导和基因表达中的调节作用，为通过操纵NOC/OFQ和ORL1功能的新策略的基础，通过操纵GAIP（RGS）基因表达和活动，因此可以在疼痛和其他方面进行治疗和其他neurorders和其他neurorders的策略。