Dynamics of translation under normal conditions and oxidative stress

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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.