Dynamics of translation under normal conditions and oxidative stress

正常条件和氧化应激下的翻译动力学

• 批准号: 220072946
• 负责人: Professorin Dr. Zoya Ignatova
• 金额: --
• 依托单位: Institut für Biochemie und Molekularbiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/220072946?language=en
• 关键词: Dynamics; translation; under; normal; conditions

Ribosomes transiently attenuate their progress along the mRNAs and this non-uniform speed plays an important role in gene expression and protein biogenesis, including co-translational folding and protein targeting. The effect of external stress on the translation dynamics and attenuation remains enigmatic. In this project, we seek to assess the integrated response of the cell on the level of translation upon exposure to oxidative stress. We will use ribosome profiling combined with mRNA sequencing to obtain a global, cell-wide view of translated mRNAs with single codon resolution accuracy. Comparison between cells grown in normal balanced conditions with those exposed to oxidative agents will reveal the programs operating in cells to counteract oxidative stress.Stress granules (SG) and P-bodies are part of the integrated stress response in which cytosolic and nuclear proteins temporarily sequester mRNAs and withdraw them from translation. In parallel, to the global assessment of the genes that shape the cellular response to oxidative stress, we aim to determine the mRNAome of stress granules and P-bodies using the power of deep sequencing approaches. Furthermore, we will address the dynamics of mRNA distribution between these RNA particles and translating ribosomes upon exposure to oxidative stress using selective pulse-labeling approaches combined with immunoprocipitation. Together this data will provide a comprehensive global view on the dynamics of the translation under stress exposure and will reveal the effect of translation in modulating cellular stress response.

核糖体瞬时减弱它们沿着mRNA的进展，并且这种不均匀的速度在基因表达和蛋白质生物合成中起重要作用，包括共翻译折叠和蛋白质靶向。外部应力对平移动力学和衰减的影响仍然是个谜。在这个项目中，我们试图评估暴露于氧化应激后细胞在翻译水平上的综合反应。我们将使用核糖体分析与mRNA测序相结合，以获得具有单密码子解析准确性的翻译mRNA的全局，细胞范围的视图。比较在正常平衡条件下生长的细胞与暴露于氧化剂的细胞将揭示细胞中对抗氧化应激的程序。应激颗粒（SG）和P体是整合的应激反应的一部分，其中胞质和核蛋白暂时隔离mRNA并使其从翻译中撤回。与此同时，为了对塑造细胞对氧化应激反应的基因进行全球评估，我们的目标是使用深度测序方法的力量来确定应激颗粒和P体的mRNA组。此外，我们将解决这些RNA颗粒和翻译核糖体暴露于氧化应激后，使用选择性脉冲标记结合免疫结合的方法之间的mRNA分布的动态。总之，这些数据将提供一个全面的全球视野下的动态翻译应激暴露，并将揭示翻译在调节细胞应激反应的影响。