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：
8628848
负责人：
Elizabeth J Tran
金额：
$ 28.2万
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 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8628848
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 Functional RNA Gene Expression Gene Expression Profile Gene Expression Regulation Gene Proteins Genes Genetic Genetic Transcription 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 Work Yeasts base biological research cancer type cell growth chromatin remodeling driving force gene repression 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 public health relevance 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，精心策划的过程之一。大量研究表明，诸如转录，信使RNA（mRNA）加工和mRNA衰变之类的步骤是高度耦合的，形成了用于调节蛋白质编码基因的复杂分子回路。此过程的核心是一类称为死盒蛋白的RNA解旋酶，它们在RNA生物学的各个方面都起着重要作用。 DBP2是在酿酒酵母萌芽的酵母菌中，在很大程度上未表征的Dead-box RNA解旋酶家族。尽管许多研究表明，DBP2，HDDX5或P68的人（H）直系同源物作为转录调节剂，但在此过程中尚未描述DBP2的转录作用，HDDX5的精确分子功能尚不清楚。我们的研究现在提供了第一个证明，即在萌芽酵母中核基因表达步骤需要DBP2，在mRNA生物发生和染色质重塑之间的界面上起作用。此外，我们的工作表明DBP2在RNA聚合酶II的非蛋白质编码RNA合成的位点起作用。本文中，我们建议使用生化，分子和遗传方法的组合获得对DBP2的详细理解。在AIM I中，我们将利用一系列体外测定法来生化表征DBP2，然后分析酶促的正常细胞生长和基因表达的需求。在AIM II中，我们将通过分子和遗传方法的结合来确定DBP2如何在基因表达中起作用。在AIM III中，我们将定义和表征体内分子相互作用，这些相互作用使DBP2成为“感觉”新生的转录本。我们的长期工作模型是DBP2是调节转录组的基因表达电路中的一种酶“切换”。因此，这些研究有可能揭示全基因组表观遗传调节的新机制，这是一项反映生物医学研究所面临的当前挑战的NIH战略计划。重要的是，许多死盒蛋白基因与包括癌症，神经系统疾病和艾滋病在内的人类疾病状态有关。因此，发现单个死盒蛋白的生理作用是基础生物学研究和医学界的主要挑战。