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（由申请人提供）：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对调节细胞功能的信号作出反应。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决定，这是一个基本的加工步骤，涉及在polyA（pA）位点切割mRNA前体，然后将腺苷添加到新的3'端。改变pA位点可以改变编码序列的量或去除3'非翻译区（UTR）中控制定位、翻译和稳定性的重要调控序列。 大多数真核基因含有两个或多个PA位点，其产生不同长度的mRNA同种型。在发育、分化和肿瘤发生过程中，这些亚型的比例发生了惊人的变化，并对细胞的环境作出反应。这些发现表明，选择性多聚腺苷酸化（阿帕）加入转录起始和选择性剪接作为一个重要的，但未得到充分重视的方式来调节特定细胞状态所需的mRNA的数量和类型。 然而，导致阿帕的机制还没有很好地定义，对蛋白质输出的后果和对细胞功能的贡献仍然知之甚少。我们的总体目标是了解PA网站使用这些广泛的变化的机制和功能后果。该领域以前的工作主要集中在3 'UTR的变化，以及由于调控序列的获得或丢失而引起的转录后调控的变化。与此相反，相对较少的关注已给予结束在编码序列（CDS-PA）内的PA网站的成绩单，尽管有明确的证据表明，他们的患病率和系统的变化，在细胞状态的变化。因此，我们将把我们的研究重点放在CDS-pA转录本上。我们将测试的假设，这些异构体的平衡的变化是由mRNA的稳定性，mRNA的多聚腺苷酸化，转录终止的变化的协调组合控制，无论机制，这些变化将影响蛋白质的生产，因此是一个适当的响应环境变化的组成部分。我们的方法是创新的，因为它将使用全基因组表达分析来指导集中的分子生物学实验，这些实验将探测基因表达调控中重要步骤的潜在机制。这一建议是重要的，因为阿帕的广泛使用及其迅速影响细胞蛋白质的数量和类型的潜力。这些目标的实现，预计将产生新的见解，广泛适用于其他细胞状态调制的替代聚腺苷酸化。