Chromatin Dynamics of the CFTR locus

CFTR 位点的染色质动力学

基本信息

批准号：
8279234
负责人：
Martin John Walsh
金额：
$ 42.38万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8279234
关键词：
Achievement Alleles Architecture Binding Biochemical Biological Assay Cell Nucleus Cells ChIP-seq Characteristics Chromatin Chromatin Structure Chromosome Structures Clinical Complex Coupled Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator DNA DNA-Binding Proteins Development Disease Disease Outcome Embryo Enhancers Environment Epigenetic Process Euchromatin Family Foundations Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Genetic Polymorphism Genetic Recombination Genetic Transcription Genome Genomics Goals Health Hereditary Disease Heterochromatin Histone Acetylation Histones Human Human Genetics Individual Knowledge Laboratories Location Mediating Methylation Modeling Molecular Molecular Conformation Molecular Genetics Mus Mutation Nuclear Nuclear Protein Nuclear Proteins Nucleosomes Occupations Pancreas Patients Pattern Phenotype Physiology Post-Translational Protein Processing Protein Complex Subunit Protein Family Proteins Quality of life RNA RNA Polymerase I RNA Polymerase II Regulation Regulator Genes Reporting Role Shapes Site Stress Tertiary Protein Structure Therapeutic Transcriptional Regulation Transgenic Organisms Variant Work base chromatin remodeling cystic fibrosis patients design gene discovery helicase histone modification homologous recombination in vivo member preimplantation programs promoter protein function scaffold

项目摘要

DESCRIPTION (provided by applicant): Since the discovery of the gene responsible for cystic fibrosis (CF) encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein, several therapeutic approaches have emerged that have extended and enhanced the quality of life for patients with CF. However, such advances in clinical approaches used for treating CF still remain at odds with our current knowledge involving the epigenetic regulation and transcriptional control of the CFTR gene (CFTR). Despite the molecular and genetic characterization of mutations in patients and families afflicted with CF, the relationship of well -characterized polymorphisms still remain inconsistent in predicting disease outcome associated with CF. This reality stresses a lack in understanding how CFTR is regulated and how transcription of corresponds with the CF phenotype. The objective of this application is to determine the fundamental mechanisms in chromatin that regulate CFTR transcription. Gene regulation occurs within the native chromatin environment through the alteration of nucleosome mobility, histone content or variation, and post-translational modification of histones in chromatin. Furthermore, less is still known of the three dimensional (3D) characteristics of CFTR transcription requiring the cooperative interactions between other gene loci that give rise to the high order chromosomal organization for temporal and spatial control. Despite the advance in the characterizing many covalent post-translational histone modifications and their contribution to the epigenetic programming of gene expression, absent among this particular scientific milestone has been the functional characterization of ATP - dependent chromo - helicase domain (CHD) proteins and their contribution to specific gene expression programs. Recent genetic and biochemical examination of only a few mammalian CHD proteins reveal the remarkable importance of these gene products during development and in disease. Therefore, efforts to functionally characterize the individual members of the CHD family of proteins will likely have profound impact on human health and understanding disease patho-physiology. We propose the following specific aims. First, we will investigate the role of CHD6 to cooperate with additional transcriptional proteins such as CTCF to regulate CFTR. These studies will characterize specific nuclear protein interactions with CHD6 to regulate CFTR gene transcription. Second, we will determine the patho -physiology associated with conditional disruption of CHD6 in mice and study the consequence on the epigenetic signature and transcription of CFTR. Finally, we propose to characterize the role of CHD6 to participate as a stricture with CTCF for the convergence of multiple loci with CFTR to coordinate a 3D transcriptional program using ChIP sequencing, 3C/4C, and RNA FISH approaches to understand the modified or variant histone signatures and dynamics of chromosomal exchange associated with CHD6.

描述（由申请人提供）：自从发现编码囊性纤维化跨膜电导调节蛋白（CFTR）蛋白的囊性纤维化（CF）基因以来，已经出现了几种治疗方法，延长和提高了 CF 患者的生活质量。然而，用于治疗 CF 的临床方法的此类进展仍然与我们目前涉及 CFTR 基因 (CFTR) 表观遗传调控和转录控制的知识不一致。尽管患有 CF 的患者和家庭的突变具有分子和遗传特征，但充分表征的多态性在预测与 CF 相关的疾病结果方面仍然不一致。这一现实强调缺乏对 CFTR 如何调节以及 CFTR 的转录如何与 CF 表型相对应的了解。本应用的目的是确定染色质中调节 CFTR 转录的基本机制。基因调控通过改变核小体迁移性、组蛋白含量或变异以及染色质中组蛋白的翻译后修饰而发生在天然染色质环境中。此外，人们对 CFTR 转录的三维 (3D) 特征知之甚少，这些特征需要其他基因位点之间的协作相互作用，从而产生用于时间和空间控制的高阶染色体组织。尽管在表征许多共价翻译后组蛋白修饰及其对基因表达的表观遗传编程的贡献方面取得了进展，但在这一特定的科学里程碑中缺少 ATP 依赖性染色解旋酶结构域 (CHD) 蛋白的功能表征及其贡献特定的基因表达程序。最近仅对少数哺乳动物先心病蛋白进行的遗传和生化检查揭示了这些基因产物在发育和疾病过程中的显着重要性。因此，对 CHD 蛋白质家族各个成员进行功能表征的努力可能会对人类健康和理解疾病病理生理学产生深远的影响。我们提出以下具体目标。首先，我们将研究CHD6与CTCF等其他转录蛋白合作调节CFTR的作用。这些研究将表征特定核蛋白与 CHD6 的相互作用，以调节 CFTR 基因转录。其次，我们将确定小鼠中与条件性破坏 CHD6 相关的病理生理学，并研究其对表观遗传特征和 CFTR 转录的影响。最后，我们建议描述 CHD6 作为 CTCF 的限制参与多个位点与 CFTR 的融合的作用，以协调使用 ChIP 测序、3C/4C 和 RNA FISH 方法的 3D 转录程序来了解修饰或变异的组蛋白与 CHD6 相关的染色体交换的特征和动态。