Effect of oxidative stress on the association of long noncoding RNA and mRNA with ribosomes

In this study we analysed the association of long noncoding RNAs (lncRNAs) and mRNAs with ribosomes in control and oxidatively stressed (2hr treatment with H2O2) MRC5 fibroblasts. A custom microarray was used to analyze the stress-induced distribution changes of transcripts between the non-translating (free RNP and 80S), low-translating (two, three and four ribosomes), and highly-translating (five and more ribosomes) pools in three experiments. The finding suggests that stress-induced lncRNAs showed a higher polysome occupancy as compared to the transcripts of coding genes in response to oxidative stress. Samples from 23 polysome fractions were divided into 3 pools: untranslated (fractions 1-9,representing free RNPs and 80S ribosomes); low-translated (fractions 10-14, representing 2-4 ribosomes) and highly-translated (fractions 15-23 representing 5 and more ribosomes). The procedure was carried out three times, each with an independent RNA preparation and each analyzed with 3 microarrays, both in control and OS condition (2 hours) together with whole-cell RNA for each condition.