Dynamics of translation under normal conditions and oxidative stress

核糖体会短暂地减弱其沿着mrna的进程，这种不均匀的速度在基因表达和蛋白质生物发生中起着重要作用，包括共翻译折叠和蛋白质靶向。外应力对平移动力学和衰减的影响仍然是一个谜。在这个项目中，我们试图评估细胞在暴露于氧化应激的翻译水平上的综合反应。我们将使用核糖体分析结合mRNA测序来获得具有单密码子分辨率准确性的翻译mRNA的全局，细胞范围视图。比较在正常平衡条件下生长的细胞和那些暴露于氧化剂的细胞，将揭示细胞中运行的程序来抵消氧化应激。应激颗粒（SG）和p -小体是综合应激反应的一部分，其中细胞质和核蛋白暂时隔离mrna并使其退出翻译。与此同时，为了全面评估影响细胞对氧化应激反应的基因，我们的目标是利用深度测序方法确定应激颗粒和p体的mRNAome。此外，我们将利用选择性脉冲标记方法结合免疫沉淀，解决暴露于氧化应激下这些RNA颗粒和翻译核糖体之间mRNA分布的动力学问题。总之，这些数据将提供一个全面的视角，在压力暴露下翻译的动态，并将揭示翻译在调节细胞应激反应中的作用。

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.