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：
8827802
负责人：
Elizabeth J Tran
金额：
$ 28.08万
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 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8827802
关键词：
ATP phosphohydrolase Acquired Immunodeficiency Syndrome Binding Binding Sites Biochemical Biochemical Pathway Biogenesis Biological Assay Biological Models Biology Biomedical Research Boxing Cells Code Communities Complex Coupled Enzymes Epigenetic Process Eukaryota Family Gene Expression Gene Expression Profile Gene Expression Regulation Gene Proteins Genes Genetic Genetic Transcription Health Human In Vitro Individual Knowledge Laboratories Life Link Malignant Neoplasms Mammalian Cell Maps Mediating Medical Messenger RNA Modeling Molecular Molecular Chaperones Molecular Genetics Normal Cell Nuclear Oncogenes Organism Orthologous Gene Physiological Play Process Protein Family Proteins RNA RNA Binding RNA Decay RNA Helicase RNA Polymerase II RNA Processing RNA chemical synthesis Regulation Repression Resistance Role Saccharomyces Saccharomyces cerevisiae Saccharomycetales Series Site Transcript United States National Institutes of Health Untranslated RNA Work Yeasts base biological research cancer type cell growth chromatin remodeling driving force epigenetic regulation gene repression genetic approach genome-wide helicase human disease in vitro Assay in vitro activity in vivo insight member nervous system disorder novel overexpression protein complex protein protein interaction transcription factor tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Gene expression in eukaryotes is one of the most complex, highly orchestrated processes in living organisms. Numerous studies have shown that steps such as transcription, messenger RNA (mRNA) processing and mRNA decay, are highly coupled, forming an intricate molecular circuit for regulation of protein-coding genes. Central to this process, are a class of RNA helicases called DEAD-box proteins that perform essential roles in all aspects of RNA biology. Dbp2 is a largely uncharacterized member of the DEAD-box RNA helicase family in the budding yeast Saccharomyces cerevisiae. Whereas numerous studies have shown that the human (h) ortholog of Dbp2, hDDX5 or p68, functions as a transcriptional regulator, no transcriptional role has been described for Dbp2 and the precise molecular function of hDDX5 in this process is not understood. Our studies now provide the first demonstration that Dbp2 is required for nuclear gene expression steps in budding yeast, functioning at the interface between mRNA biogenesis and chromatin remodeling. Furthermore, our work demonstrates that Dbp2 functions at sites of non-protein coding RNA synthesis by RNA polymerase II. Herein, we propose to obtain a detailed understanding of Dbp2 using a combination of biochemical, molecular and genetic approaches. In Aim I, we will utilize a series of in vitro assays to biochemically characterize Dbp2 and subsequently analyze the enzymatic requirements for normal cell growth and gene expression. In Aim II, we will determine how Dbp2 functions in gene expression through a combination of molecular and genetic approaches. In Aim III, we will define and characterize the in vivo molecular interactions that enable Dbp2 to 'sense' nascent transcripts. Our long term working model is that Dbp2 is an enzymatic 'toggle' in the gene expression circuit that regulates the transcriptome. Thus, these studies have the potential to reveal novel mechanisms for genome-wide epigenetic regulation, an NIH strategic initiative reflective of the current challenges facing biomedical research. Importantly, numerous DEAD-box protein genes have been linked to human disease states including cancer, neurological disorders and AIDS. Therefore, uncovering the physiological role of individual DEAD-box proteins is a major challenge to basic biological research and the medical community.

描述（由申请人提供）：真核生物中的基因表达是生物体中最复杂、高度协调的过程之一。大量研究表明，转录、信使 RNA (mRNA) 加工和 mRNA 衰变等步骤高度耦合，形成了调控蛋白质编码基因的复杂分子回路。这一过程的核心是一类称为 DEAD-box 蛋白的 RNA 解旋酶，它们在 RNA 生物学的各个方面发挥着重要作用。 Dbp2 是芽殖酵母酿酒酵母中 DEAD-box RNA 解旋酶家族的一个很大程度上未被表征的成员。尽管大量研究表明 Dbp2、hDDX5 或 p68 的人类 (h) 直系同源物具有转录调节因子的功能，但尚未描述 Dbp2 的转录作用，并且 hDDX5 在此过程中的精确分子功能尚不清楚。我们的研究现在首次证明 Dbp2 是芽殖酵母核基因表达步骤所必需的，在 mRNA 生物发生和染色质重塑之间的界面发挥作用。此外，我们的工作表明，Dbp2 在 RNA 聚合酶 II 合成非蛋白质编码 RNA 的位点发挥作用。在此，我们建议结合生化、分子和遗传学方法来详细了解 Dbp2。在目标 I 中，我们将利用一系列体外测定来对 Dbp2 进行生化表征，并随后分析正常细胞生长和基因表达的酶需求。在目标 II 中，我们将通过分子和遗传学方法的结合来确定 Dbp2 在基因表达中的功能。在目标 III 中，我们将定义和表征使 Dbp2 能够“感知”新生转录本的体内分子相互作用。我们的长期工作模型是 Dbp2 是基因表达回路中调节转录组的酶“开关”。因此，这些研究有可能揭示全基因组表观遗传调控的新机制，这是 NIH 的一项战略举措，反映了生物医学研究当前面临的挑战。重要的是，许多死亡盒蛋白基因与人类疾病状态有关，包括癌症、神经系统疾病和艾滋病。因此，揭示单个DEAD-box蛋白的生理作用是基础生物学研究和医学界面临的重大挑战。