Analysis of HIF and mTOR dependent translational regulation of mRNAs resolved by their transcription start sites, using HP5 technology

Changes in translational efficiency of mRNAs consequent on alteration of HIF and/or mTOR pathway activity in VHL-defective kidney cancer cell were analysed. High-resolution polysome profiling followed by sequencing of the 5' ends of mRNAs (HP5) was used to measure translational efficiency of mRNAs resolved by their transcription start sites. HP5 was applied to VHL-defective kidney cancer cell lines with or without VHL reintroduction and/or mTOR inhibition with Torin 1 treatment. In addition, the contribution of HIF2A/EIF4E2 pathway was assessed using 786-O cells in which EIF4E2 was inactivated.