Nuclear Pore Complexes As Scaffolds For Genome Architecture And Epigenetic Maintenance

核孔复合物作为基因组结构和表观遗传维护的支架

基本信息

批准号：
10701898
负责人：
Maya Capelson
金额：
$ 32.55万
依托单位：
SAN DIEGO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10701898
关键词：
Affect Architecture Binding Biology Cell Nucleus Cells Chromatin Chromatin Structure Complex Cytoplasm DNA Deposition Development Developmental Gene Developmental Process Distant Drosophila genome Ecdysone Elements Enhancers Environment Epigenetic Process Event Exposure to Future Gene Activation Gene Expression Gene Expression Regulation Gene Silencing Genes Genetic Transcription Genome Genomics Goals Heart Diseases Hematologic Neoplasms Histones Homeobox Genes Hormones Human Pathology Immunoprecipitation Investigation Knowledge Link Macromolecular Complexes Maintenance Mediating Memory Methods Molecular Nuclear Nuclear Envelope Nuclear Matrix Nuclear Pore Nuclear Pore Complex Nuclear Pore Complex Proteins Pathology processes Pathway interactions Phenotype Play Polycomb Proteins Regulation Role Testing Tissues Transcriptional Activation Transcriptional Regulation chromatin immunoprecipitation chromosome conformation capture driving force embryonic cell culture epigenetic memory experimental study fly genome-wide nucleocytoplasmic transport promoter recruit response scaffold

项目摘要

The Nuclear Pore Complex (NPC) is a nuclear envelope embedded multi-component complex, which mediates transport of molecules between the nucleus and the cytoplasm. In addition to their classical function in transport, NPC components (Nups) have been implicated in transcriptional regulation via binding to the genome. Yet what regulatory steps of transcription are controlled by Nups and how chromatin-binding roles of Nups contribute to metazoan development is currently unclear. We identified the binding of multiple Nups to hundreds of promoters and enhancers in the Drosophila genome, and discovered a previously unreported role of the NPC in the formation of enhancer-promoter loops. Specifically, we identified Nup98 to be required for the formation of an enhancer-promoter loop at a gene activated by a developmental hormone ecdysone. Functionally, we found that the loss of Nup98-mediated enhancer-promoter loop affected the primed response to subsequent activation or transcriptional memory. Interestingly, ecdysone-regulated genes stably associated with nuclear pores before and after activation, suggesting that metazoan NPCs can be utilized as an organizing scaffold for genes awaiting future activation events. Together, these findings implicate Nups as a new class of architectural proteins for enhancers and suggest that stabilization of enhancer-promoter loops by nuclear pore binding constitutes a mechanism of epigenetic maintenance. We project that this function of the nuclear pore will be highly relevant to gene regulation during metazoan development. To be able to investigate the genome- organizing role of Nups in developmental gene regulation, we first plan to identify molecular determinants of Nup-mediated enhancer-promoter looping. Thus, in Aim 1, we will define which Nups and which other architectural proteins participate in the establishment of ecdysone- induced genomic loops. Additionally, we plan to identify DNA elements that are sufficient to tether to the NPC or that are necessary for loop stabilization. In Aim 2, we propose to define the relationship between formation of Nup-mediated enhancer-promoter loops and transcriptional activation and memory, by identifying chromatin changes that occur as a specific consequence of looping. Furthermore, we will examine the effect of Nups on maintenance of genomic loops in fly tissues during development and identify a comprehensive set of genomic contacts that are regulated by Nups. Together, these experiments are expected to expand our knowledge of the driving forces and principles of genome architecture, gene expression and nuclear pore biology.

核孔复合物（NPC）是一个核包膜嵌入的多组分复合物，该复合物介导细胞核和分子的转运细胞质。除了它们在运输方面的经典功能外，NPC组件（NUP）已经通过与基因组结合进行转录调节。然而，什么监管步骤转录受NUP的控制以及NUP的染色质结合作用如何促进后生开发目前尚不清楚。我们确定了多个NUP与数百个启动子的结合以及果蝇基因组中的增强剂，并发现了NPC先前未报告的作用增强剂启动器循环的形成。具体来说，我们确定了NUP98是必需的在由发育激素ecdysone激活的基因上形成增强子启动器循环。在功能上，我们发现NUP98介导的增强剂启动器的损失影响了Primed 对随后的激活或转录内存的响应。有趣的是，ecdysone调节的基因激活前后与核孔相关，表明后生NPC可以被用作等待未来激活事件的基因的组织脚手架。在一起，这些调查结果暗示NUP是增强剂的新型建筑蛋白质，并建议通过核孔结合对增强子启动器环的稳定构成了表观遗传学的机制维护。我们预计核孔的这种功能将与基因调节高度相关在多宗开发期间。能够研究NUP在发育基因调节，我们首先计划鉴定NUP介导的分子决定因素增强器促销循环。因此，在AIM 1中，我们将定义哪些NUP和哪些其他建筑蛋白质参与依dysone诱导的基因组环的建立。此外，我们计划识别足以将其绑定到NPC或循环所需的DNA元素稳定。在AIM 2中，我们建议定义NUP介导的形成之间的关系通过识别染色质的变化，增强剂促销循环以及转录激活和内存这是循环的特定结果。此外，我们将研究NUPS对在开发过程中维持蝇组织中的基因组环，并确定一组综合由NUPS调节的基因组接触。共同期望这些实验扩大我们的了解基因组结构，基因表达和核原理的知识毛孔生物学。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Nuclear pores in genome architecture and enhancer function.

DOI：
10.1016/j.ceb.2019.04.001
发表时间：
2019-06
期刊：
Current opinion in cell biology
影响因子：
7.5
作者：
Pau Pascual-Garcia;M. Capelson
通讯作者：
Pau Pascual-Garcia;M. Capelson

Metazoan Nuclear Pores Provide a Scaffold for Poised Genes and Mediate Induced Enhancer-Promoter Contacts.

DOI：
10.1016/j.molcel.2017.02.020
发表时间：
2017-04-06
期刊：
Molecular cell
影响因子：
16
作者：
Pascual-Garcia P;Debo B;Aleman JR;Talamas JA;Lan Y;Nguyen NH;Won KJ;Capelson M
通讯作者：
Capelson M

The nuclear pore complex and the genome: organizing and regulatory principles.

DOI：
10.1016/j.gde.2021.01.005
发表时间：
2021-04
期刊：
Current opinion in genetics & development
影响因子：
4
作者：
Pascual-Garcia P;Capelson M
通讯作者：
Capelson M

Preparation of Drosophila Polytene Chromosomes, Followed by Immunofluorescence Analysis of Chromatin Structure by Multi-fluorescence Correlations.

果蝇多线染色体的制备，然后通过多荧光相关性对染色质结构进行免疫荧光分析。

DOI：
10.21769/bioprotoc.3673
发表时间：
2020
期刊：
Bio-protocol
影响因子：
0.8
作者：
Kuhn,TerraM;Little,ShawnC;Capelson,Maya
通讯作者：
Capelson,Maya

Correct dosage of X chromosome transcription is controlled by a nuclear pore component.

DOI：
10.1016/j.celrep.2021.109236
发表时间：
2021-06-15
期刊：
Cell reports
影响因子：
8.8
作者：
Aleman JR;Kuhn TM;Pascual-Garcia P;Gospocic J;Lan Y;Bonasio R;Little SC;Capelson M
通讯作者：
Capelson M