Regulation of eukaryotic mRNA polyadenylation by sustained stress

持续应激对真核 mRNA 多腺苷酸化的调节

• 批准号: 9055727
• 负责人: CLAIRE L MOORE
• 金额: $ 32.57万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9055727
• 关键词: 3' Untranslated Regions; Adenosine; Affect; Alternative Splicing; Attention; Bioinformatics; Biological; Carbon; Cell Extracts; Cell physiology; Cells; Code; Coupling; Data Set; Defect; Degradation Pathway; Development; Disease; Environment; Equilibrium; Eukaryotic Cell; Exhibits; Fermentation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genetic screening method; Glucose; Glycerol; Growth; Health; In Vitro; Initiator Codon; Knowledge; Lead; Length; Location; Malignant Neoplasms; Maps; Messenger RNA; Metabolism; Mission; Mitochondria; Molecular; Normal Cell; Output; Pathway interactions; Pattern; Polyadenylation; Polyadenylation Pathway; Post-Transcriptional Regulation; Prevalence; Process; Production; Protein Isoforms; Proteins; Regulation; Research; Respiration; Role; Saccharomyces cerevisiae; Signal Transduction; Site; Source; Stress; Terminator Codon; Testing; Trans-Activators; Transcript; Transcription Initiation; Translations; Variant; Work; Yeasts; base; environmental change; experience; falls; genome integrity; human disease; innovation; insight; mRNA Precursor; mRNA Stability; mutant; novel; promoter; research study; response; transcription termination; treatment strategy; tumorigenesis; whole genome

 DESCRIPTION (provided by applicant): Eukaryotic cells often need to modify their gene expression patterns to adapt to environmental changes or to respond to signals regulating cell function. One rapid means of regulating expression is to vary the length of mRNAs. Length at the 3' end is determined by polyadenylation, an essential processing step involving cleavage of mRNA precursor at the polyA (pA) site, followed by addition of adenosines to the new 3' end. Changing the pA site can alter the amount of coding sequence or remove important regulatory sequences in the 3' untranslated region (UTR) that govern localization, translation, and stability. The majority of eukaryotic genes contain two or more pA sites which give mRNA isoforms of different length. Changes in the proportion of these isoforms occurs for a surprisingly large number of genes during development, differentiation, and tumorigenesis and in response to the cell's environment. These findings indicate that alternative polyadenylation (APA) joins transcription initiation and alternative splicing as an important, but under-appreciated way to modulate the amount and types of mRNAs needed for specific cellular states. However, the mechanisms leading to APA are not well defined, and the consequences on protein output and contribution to cell function remain poorly understood. Our overall objective is to understand the mechanism and functional consequences of these widespread changes in pA site usage. Previous work in the field has largely focused on changes to the 3'UTR, and the consequent changes in post-transcriptional regulation due to gain or loss of regulatory sequences. In contrast, relatively little attention has been given to transcripts ending at pA sites within the coding sequence (CDS-pA), despite clear evidence of their prevalence and their systematic variation in response to changes in cell state. We will therefore focus our studies on the CDS-pA transcripts. We will test the hypothesis that variations in the balance of these isoforms are controlled by a coordinated combination of changes in mRNA stability, mRNA polyadenylation, and transcription termination and that regardless of the mechanism, these changes will affect protein production and therefore are an integral component of an appropriate response to environmental changes. Our approach is innovative because it will use whole genome expression analysis to guide focused molecular biological experiments that will probe mechanisms underlying an important step in regulation of gene expression. This proposal is significant because of the wide-spread use of APA and its potential to rapidly affect the amount and type of protein made by a cell. Accomplishment of these aims is expected to yield novel insights broadly applicable to other cellular states modulated by alternative polyadenylation.

 描述（由应用程序提供）：真核细胞通常需要修改其基因表达模式以适应环境变化或响应信号调节细胞功能。调节表达的一种快速方法是改变mRNA的长度。 3'端的长度由聚腺苷酸化确定，聚腺苷酸化是一个必不可少的处理步骤，涉及在Polya（PA）位点裂解mRNA前体，然后将腺苷添加到新的3'端。更改PA位点可以更改编码序列的量，也可以在3'非翻译区域（UTR）中删除主管定位，翻译和稳定性的重要调节序列。 大多数真核基因包含两个或多个PA位点，这些位点具有不同的长度mRNA同工型。在发育，分化和肿瘤发生过程中，这些同工型的比例发生变化以及对细胞环境的响应发生的大量基因。这些发现表明，替代聚腺苷酸化（APA）将转录引发和替代剪接作为一种重要但不受欢迎的方法来调节特定细胞状态所需的mRNA的数量和类型。但是，导致APA的机制尚未很好地定义，并且对蛋白质输出的后果以及对细胞功能的贡献仍然很少了解。我们的总体目标是了解PA站点使用中这些宽度变化的机制和功能后果。该领域的先前工作主要集中在3'UTR的变化上，以及由于调节序列的增益或损失而导致的转录后调节后果。相比之下，对在编码序列（CDS-PA）内的PA位点结束的转录本的关注很少，目的地清晰的证据证明了其患病率和对细胞状态变化的响应响应。因此，我们将把研究重点放在CDS-PA转录本上。我们将测试以下假设：这些同工型平衡的变化是由mRNA稳定性，mRNA聚腺苷酸化和转录终止的协调组合控制的，无论机制如何，这些变化都会影响蛋白质的产生，因此是对环境变化的适当响应的整体组成部分。我们的方法具有创新性，因为它将使用整个基因组表达分析来指导聚焦的分子生物学实验，该实验将探测调节基因表达的重要步骤的基础机制。该提议很重要，因为APA的广泛使用及其潜力快速影响细胞制造的蛋白质的数量和类型。这些目标的实现有望产生广泛适用于其他通过替代聚腺苷化调节的其他细胞状态的新颖见解。