Dynamics of translation under normal conditions and oxidative stress

核糖体短暂地减弱了它们沿着mRNAs的进展，这种不均匀的速度在基因表达和蛋白质生物发生中发挥着重要作用，包括共翻译折叠和蛋白质靶向。外界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，我们试图评估细胞在氧化应激下在翻译水平上的综合反应。我们将使用核糖体图谱和信使核糖核酸测序相结合的方法，获得具有单一密码子解析精度的翻译的mRNAs的全局性、细胞范围的视图。将在正常平衡条件下生长的细胞与暴露在氧化剂中的细胞进行比较，将揭示在细胞中运行的抵消氧化应激的程序。应激颗粒(SG)和P-小体是综合应激反应的一部分，在该反应中，胞浆和核蛋白暂时隔离mRNAs并将其从翻译中撤回。同时，在对影响细胞对氧化应激反应的基因进行全球评估的同时，我们的目标是利用深度测序方法的力量来确定应激颗粒和P-体的mRNAome。此外，我们将使用选择性脉冲标记方法和免疫沉淀相结合的方法，研究氧化应激下这些RNA颗粒和翻译核糖体之间的mRNA分布的动态。总而言之，这些数据将为翻译在压力下的动态提供一个全面的全球视角，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.