Regulation of Mutationally Activated Gq/11

突变激活 Gq/11 的调控

基本信息

批准号：
10376872
负责人：
PHILIP B WEDEGAERTNER
金额：
$ 31.2万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10376872
关键词：
Address Adult Binding Biochemical Biological Assay Biosensor Blindness Blood capillaries Cardiovascular Physiology Cell Differentiation process Cell membrane Cell model Cell physiology Cells Codon Nucleotides Congenital Hemangioma Cultured Cells Cyclodepsipeptides Data Disease Endocrine System Diseases Fluorescence Microscopy G-Protein-Coupled Receptors GNAQ gene GTP Binding GTP-Binding Proteins Genes Glutamine Guanine Nucleotide Exchange Factors Guanosine Triphosphate Heart Diseases Heterotrimeric G Protein Subunit Heterotrimeric GTP-Binding Proteins Hydrolysis Hypertension Immune Immunofluorescence Microscopy Knowledge Link Location Malignant Neoplasms Mediating Membrane Mitogen-Activated Protein Kinases Modeling Molecular Mutation Mutation Analysis Natural Products Neoplasm Metastasis Neoplasms in Vascular Tissue Neurotransmitters Oncogenic Pathway interactions Patients Pharmacology Phospholipase C Physiological Physiological Processes Plasma Cells Play Protein Subunits Proteins Refractory Regulation Research Project Grants Research Proposals Resistance Role Signal Pathway Signal Transduction Smell Perception Somatic Mutation Sturge-Weber Syndrome Surface Taste Perception Testing Uveal Melanoma Vision Work base cell growth regulation cell motility disease-causing mutation experience extracellular genetic regulatory protein human disease inhibitor live cell microscopy malformation malignant neoplasm of eye mutant nervous system disorder neuron development new therapeutic target novel palmitoylation protein function receptor-mediated signaling reconstitution resistance mechanism response rho therapeutic target trafficking

项目摘要

Abstract Heterotrimeric G proteins () are well known for their function in linking G protein-coupled receptors (GPCRs) to a variety of intracellular responses, and thereby playing essential roles in transmitting a wide variety of extracellular signals into regulation of countless physiological processes. However, it has also become increasingly recognized that G proteins can function independently of GPCRs. Highlighting the physiological relevance of GPCR-independent G protein function, constitutively activating somatic mutations in several G subunits have been identified and shown to contribute to human disease, most prominently in cancer. This research project focuses on constitutively active mutants of the highly similar q and 11 subunits, collectively termed q/11. Mutations at one of two hotspot residues, that result in inhibition of the GTP hydrolysis turn-off mechanism, can convert q/11 into an oncogenic driver of uveal melanoma, and such constitutively activating q/11 mutations have also been associated with vascular tumors, capillary malformation, congenital hemangioma and Sturge Weber syndrome. This research project will focus on uncovering basic cellular mechanisms regulating signaling by mutationally activated q/11. G subunits are typically thought as exceptionally difficult proteins for which to develop pharmacological or molecular inhibitors. However, numerous recent studies have demonstrated that the highly similar natural products YM-254890 and FR900359 are effective inhibitors of wild type and mutationally activated q/11. Our work has uncovered an unexpected mechanism of action of these inhibitors leading to the hypothesis that YM- 254890 and FR900359 inhibit q/11, at least in part, by regulating subcellular localization of q/11. Understanding the novel mechanisms of inhibition and cellular regulation of mutationally activated q/11 may suggest novel therapeutic targets or approaches for disrupting dysregulated signaling. The objectives in this research project will be pursued by a variety of experimental approaches, including using cultured cells, immunofluorescence microscopy, fluorescence microscopy of live cells, constitutive targeting to specific subcellular locations, biosensors, pharmacological inhibitors, mutational analysis, and biochemical assays.

抽象的异三聚体G蛋白（）以其在连接G蛋白偶联受体中的功能而闻名（GPCR）到各种细胞内反应，从而在传播宽阔的地方发挥着重要作用细胞外信号的多种多样，以调节无数的物理过程。但是，它也有越来越认识到G蛋白可以独立于GPCR起作用。突出显示与GPCR无关G蛋白功能的生理相关性，组成性激活体细胞突变在几个G亚基中，已被鉴定出来并证明对人类疾病有贡献，最重要的是癌症。该研究项目着重于高度相似q和11的一贯活跃的突变体亚基，共同称为Q/11。在两个热点保留之一的突变，这导致抑制 GTP水解关闭机制，可以将Q/11转换为紫veal黑色素瘤的致癌驱动器，并且这种组成症激活q/11突变也与血管肿瘤，毛细血管有关畸形，先天性血管瘤和Sturge Weber综合征。该研究项目将重点放在发现通过突变激活的Q/11调节信号传导的基本细胞机制。 G亚基是通常认为是出现药物或分子的异常困难蛋白质抑制剂。但是，最近的许多研究表明，高度相似的天然产品 YM-254890和FR900359是野生型和突变激活的Q/11的有效抑制剂。我们的工作已经发现了这些抑制剂的意外作用机理，导致假设YM- 254890和FR900359至少部分通过调节Q/11的亚细胞定位来抑制Q/11。了解突变激活的Q/11的新型抑制和细胞调节机制建议新的热目标或方法来破坏失调的信号传导。其中的对象研究项目将通过多种实验方法来追求，包括使用培养细胞，免疫荧光显微镜，活细胞的荧光显微镜，构成靶向特定的靶向亚细胞位置，生物传感器，药物抑制剂，突变分析和生化测定。