Molecular underpinnings of photoreceptor transcriptional regulation by CRX and NRL

CRX 和 NRL 光感受器转录调节的分子基础

基本信息

批准号：
10562276
负责人：
Nikolai O Artemyev
金额：
$ 38.88万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10562276
关键词：
Affinity Binding Biochemical Biological Assay Biology CREB1 gene CREB3 gene CRX protein Chromatin Complex Cone DNA DNA Binding DNA analysis DNA-Protein Interaction Defect Development Dimerization Disease Elements Exhibits FOS gene Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Transcription Glean Goals Heterodimerization Homeobox Homeodomain Proteins Homeostasis Human Individual Inherited Knock-out Knowledge Leber&apos s amaurosis Length Leucine Zippers Link Maintenance Mediating Molecular Mutation Mutation Analysis Nature Neural Retina Outcome Pathogenicity Phenotype Photoreceptors Protein Family Proteins Regulation Regulatory Element Research Response Elements Retina Retinal Cone Retinal Degeneration Retinal Diseases Retinitis Pigmentosa Rhodopsin Rod Signal Pathway Site Specific qualifier value Specificity Structure Testing Therapeutic Tissues Transcriptional Regulation Vertebrate Photoreceptors Vision Disorders Work attenuation bZIP Domain cell type cone-rod dystrophy defined contribution design disease-causing mutation follow-up genetic regulatory protein homeodomain insight mutant new therapeutic target postmitotic programs promoter protein protein interaction response retinal rods spatiotemporal synergism targeted treatment therapeutic target transcription factor treatment strategy

项目摘要

Two key transcription factors, homeodomain protein CRX and basic leucine zipper protein NRL, are at the center of gene regulation during photoreceptor differentiation and homeostasis. CRX is essential for specifying commitment of postmitotic photoreceptor precursors to the development of photoreceptor cells, whereas NRL determines the rod cell fate. In orchestrating the transcriptional program of photoreceptor development, CRX and NRL cooperate functionally and physically via a direct protein-protein interaction. Defects in photoreceptor transcriptional regulation due to mutations in the genes encoding CRX and NRL cause severe retinal diseases including retinitis pigmentosa, cone-rod dystrophy, and Leber congenital amaurosis. Despite our advanced understanding of the biology and transcriptional networks of CRX and NRL, mechanistic insight into the functions and unique synergy of these transcription factors at the atomic level is lacking. In the proposed studies, we seek to determine the crystal and solution structures of the individual DNA-bound complexes of CRX and NRL, as well as the structure of the ternary CRX/NRL/DNA complex. Although mutations in these TFs have been identified, they have not been mechanistically linked to regulation of key genes. The mechanistic predictions from the structures on how disease-causing mutations in CRX and NRL may alter DNA-binding specificity at cis-regulatory elements will be validated in the follow-up assays, including high- throughput approaches such as Spec-seq. These studies will enhance our knowledge of the functions of CRX and NRL, define the molecular nature of their synergy, and allow us to delineate specific mechanisms whereby mutant CRX and NRL proteins cause retinal diseases. We hypothesize that ultimately the structures of CRX and NRL complexed with their cis-regulatory elements will enable targeted design of therapeutics to treat visual disorders via modulation of transcriptional activities at specific promoters.

两个关键的转录因子：同源域蛋白CRX和碱性亮氨酸拉链蛋白NRL，在光感受器分化和稳态期间，位于基因调节的中心。 CRX是指定有丝分裂后感光体前体对开发的承诺至关重要光感受器细胞，而NRL决定了杆细胞的命运。在编排光感受器开发，CRX和NRL的转录程序在功能上合作以及通过直接蛋白质 - 蛋白质相互作用进行物理。光感受器转录的缺陷由于编码CRX和NRL的基因突变引起的调节引起严重的视网膜疾病包括色素性视网膜炎，锥体杆营养不良症和Leber先天性症。尽管我们对CRX和NRL的生物学和转录网络的深入了解，对这些转录因子的功能和独特协同作用的机械洞察力缺乏原子水平。在拟议的研究中，我们试图确定晶体和溶液 CRX和NRL的单个DNA结合复合物的结构，以及三元CRX/NRL/DNA复合物。尽管已经确定了这些TF中的突变，但与关键基因的调节没有机械上的联系。来自关于CRX和NRL中引起疾病突变的结构可能会改变DNA结合顺式调节元件的特异性将在后续测定中进行验证，包括高吞吐量方法，例如Spec-Seq。这些研究将增强我们对 CRX和NRL的功能，定义其协同作用的分子特性，并允许我们描绘突变CRX和NRL蛋白引起视网膜疾病的特定机制。我们假设最终CRX和NRL的结构与其顺式调节复合元素将使靶向设计的治疗剂可以通过调制来治疗视觉障碍特定启动子的转录活动。