POST-TRANSCRIPTIONAL REGULATORY COMPLEX DYNAMICS IN YEAST

酵母转录后调控复杂动态

• 批准号: 7723728
• 负责人: AIMEE M DUDLEY
• 金额: $ 1.95万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723728
• 关键词: Affect; Back; Biological Process; Cell physiology; Cells; Chimeric Proteins; Complex; Computer Retrieval of Information on Scientific Projects Database; Condition; Cytoplasmic Tail; Development; Embryonic Development; Event; Fluorescence Microscopy; Fragile X Syndrome; Funding; Gene Expression Regulation; Genes; Genetic Translation; Grant; Individual; Institution; Laboratories; Life; Localized; Mental Retardation; Messenger RNA; Neurons; Process; Proteins; RNA; RNA-Binding Proteins; Range; Research; Research Personnel; Resources; Signal Transduction; Site; Source; Transcript; Translating; Translations; United States National Institutes of Health; Work; Yeasts

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Regulation of gene expression takes place at many steps in the process of converting a gene to an active protein capable of performing its cellular function. Regulatory events that affect the stability and translation of individual mRNA transcripts are crucial for important biological processes ranging from embryogenesis (maternal RNA storage) to neuronal function (Fragile X mental retardation). However, work from many laboratories has only begun to uncover the relevant mechanistic details and environmental or developmental signals. Many important post-transcriptional regulatory events take place in dynamic cytoplasmic domains known as P-bodies. P-bodies are sites of RNA storage and degradation that can also modulate mRNA translation: RNA transcripts are translationally silenced while stored in the P-body and can be subsequently degraded or released back into the translating pool. However, the signals for determining mRNA localization to P-bodies, degradation, or release have not been identified. Using fluorescence microscopy in live yeast cells, we found that a sequence-specific RNA-binding protein, Puf3 (as a Puf3-GFP fusion), co-localizes with P-bodies (as tdimer2 RFP fusion proteins) under specific environmental conditions. This finding suggests that RNA binding proteins, like Puf3, may serve regulatory signals targeting specific sets of RNA transcripts to one or more of the processes associated with P-bodies. We hope to extend these results using fluorescence microscopy to further characterize the environmental conditions and the protein factors required for the co-localization of Puf3 and a handful of other RNA binding proteins. We would also like to examine the effects of environmental conditions and protein factors on the localization of Puf3-dependent mRNAs to P-bodies.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 基因表达的调节发生在将基因转化为能够执行其细胞功能的活性蛋白质的过程中的许多步骤。 影响单个 mRNA 转录本稳定性和翻译的调控事件对于从胚胎发生（母体 RNA 储存）到神经元功能（脆性 X 智力迟钝）等重要生物过程至关重要。 然而，许多实验室的工作才刚刚开始揭示相关的机制细节和环境或发育信号。许多重要的转录后调控事件发生在称为 P 体的动态细胞质结构域中。 P-body 是 RNA 储存和降解的位点，也可以调节 mRNA 翻译：RNA 转录物在储存在 P-body 中时翻译沉默，随后可以降解或释放回翻译池中。 然而，用于确定 mRNA 定位到 P 体、降解或释放的信号尚未确定。 在活酵母细胞中使用荧光显微镜，我们发现序列特异性 RNA 结合蛋白 Puf3（作为 Puf3-GFP 融合蛋白）在特定环境条件下与 P 体（作为 tdimer2 RFP 融合蛋白）共定位。 这一发现表明，RNA 结合蛋白（如 Puf3）可能为针对与 P 体相关的一个或多个过程的特定 RNA 转录物组提供调节信号。 我们希望使用荧光显微镜扩展这些结果，以进一步表征 Puf3 和少数其他 RNA 结合蛋白共定位所需的环境条件和蛋白质因素。 我们还想检查环境条件和蛋白质因素对 Puf3 依赖性 mRNA 定位到 P 体的影响。