The Role of DEAD-box Proteins in Gene Expression

DEAD-box 蛋白在基因表达中的作用

基本信息

批准号：
10553274
负责人：
Elizabeth J Tran
金额：
$ 47.17万
依托单位：
PURDUE UNIVERSITY
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：
2011-04-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10553274
关键词：
3&apos Untranslated Regions Address Bacteria Biochemical Biological Assay Biological Models Biological Process Biology Cells Code Cues DNA Data Defect Disease Double-Stranded RNA Enzymes Epigenetic Process Event Funding Future Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genome Genomic Instability Goals Homeostasis Human Human Pathology Hybrids In Vitro Intervention Kinetics Knowledge Laboratories Link Malignant Neoplasms Maps Metabolic Metabolic Diseases Mutation Nutrient Nutritional status Pathway interactions Process Production Proteins Publishing RNA RNA Helicase RNA Polymerase II RNA helicase A Regulation Regulator Genes Reporter Research Role Saccharomyces cerevisiae Series Structure Techniques Testing Transcript Transcriptional Regulation Untranslated RNA Virus Diseases Work Yeasts bioinformatics tool cancer type chromatin remodeling genome-wide genome-wide analysis helicase human disease in vivo innovation interdisciplinary approach malignant neurologic neoplasms member messenger ribonucleoprotein mutant nervous system disorder novel nucleic acid structure protein complex response transcription termination transcriptome sequencing

项目摘要

RNA helicases are a class of enzymes that modulate RNA structure in living cells, functioning in every aspect of RNA biology from transcription to decay. However, the precise biological function of the vast majority of the ~40 eukaryotic RNA helicases is largely unknown. In the previous funding cycle, we provided one of the first identifications of in vivo enzymatic helicase targets for a DEAD-box helicase to date. Moreover, we uncovered a novel role for RNA structure remodeling in transcription termination of RNA polymerase II in S. cerevisiae. This activity is regulated in response to environmental cues to control the expression of metabolic genes, a role that we found is conserved in human cells with the mammalian DEAD-box helicase DDX5. Strikingly, OBP2- dependent non-coding transcripts termed long non-coding RNAs (lncRNAs) form RNA-DNA hybrid structures or R-loops upon inactivation of Dbp2 that function in gene regulation. However, the mechanism(s) linking these multiple observations remains unknown. Filling this gap is of key importance because misregulation of numerous RNA helicases, including DDX5, results in disease states such as neurological disorders and cancer. Major remaining questions are: 1. How does Dbp2-dependent RNA structure impact termination? 2. What is the relationship between R-loop suppression and Dbp2? 3. Does mammalian Dbp2, termed DDX5, function in termination regulation through RNA remodeling? Our central hypothesis is that Dbp2 and DDX5 are members of a novel class of epigenetic regulators that unwind RNA structures in nascent RNAs to control transcription termination and R-loop formation in response to cellular energy status. We propose to test this hypothesis with three, focused Specific Aims, which integrate newly established and innovative strategies with proven experimental techniques. In Aim 1, we will define how RNA structure impacts transcriptional termination. In Aim 2, we will determine the mechanistic relationship between Dbp2 and R-loop suppression. In Aim 3, we will identify the enzymatic targets of DDX5 in human cells and connection to transcription termination. This research is relevant to multiple aspects of RNA biology, gene regulation, and human disease.

RNA 解旋酶是一类调节活细胞中 RNA 结构的酶，在各个方面发挥作用 RNA生物学从转录到衰变的过程。然而，绝大多数的精确生物学功能大约 40 种真核 RNA 解旋酶在很大程度上是未知的。在上一个融资周期中，我们提供了第一批迄今为止，已鉴定出 DEAD-box 解旋酶的体内酶促解旋酶靶标。此外，我们还发现 RNA 结构重塑在酿酒酵母 RNA 聚合酶 II 转录终止中的新作用。这种活动根据环境线索进行调节，以控制代谢基因的表达，这是一种作用我们发现它在人类细胞中与哺乳动物 DEAD-box 解旋酶 DDX5 是保守的。引人注目的是，OBP2- 称为长非编码 RNA (lncRNA) 的依赖性非编码转录物形成 RNA-DNA 杂合结构或 R 环在 Dbp2 失活后发挥基因调控作用。然而，连接这些的机制多项观察结果仍未知。填补这一空白至关重要，因为监管不当许多 RNA 解旋酶，包括 DDX5，会导致疾病状态，例如神经系统疾病和癌症。剩下的主要问题是： 1. Dbp2 依赖性 RNA 结构如何影响终止？ 2. R环抑制和Dbp2有什么关系？ 3. 哺乳动物 Dbp2（称为 DDX5）是否通过RNA重塑终止调节发挥作用？我们的中心假设是 Dbp2 和 DDX5 一类新型表观遗传调节因子的成员，可解开新生 RNA 中的 RNA 结构以控制响应细胞能量状态的转录终止和 R 环形成。我们建议对此进行测试具有三个重点具体目标的假设，将新建立的创新战略与经过验证的实验技术。在目标 1 中，我们将定义 RNA 结构如何影响转录终止。在目标 2 中，我们将确定 Dbp2 和 R 环抑制之间的机制关系。在目标 3 中，我们将确定人类细胞中 DDX5 的酶靶标及其与转录的联系终止。这项研究与 RNA 生物学、基因调控和人类疾病的多个方面相关。