Threonine fuels glioblastoma through YRDC-mediated codon-biased translational reprogramming (tRNA-Seq)

To understand the underlying mechanism of YRDC, an enzyme catalysing the formation of N6-threonylcarbamoyladenosine (t6A) on ANN-decoding tRNAs, and the interplay between threonine and YRDC in glioblastoma stem cells (GSCs), we carried out comparative transcriptomic analysis in patient-derived GSC456 with or without YRDC deprivation and with or without threonine restriction (control media, 800 µM threonine; threonine-restricted media, 4 µM). To further investigate the translational regulation of YRDC and threonine in GSCs, we performed ribosome profiling in GSC456 with or without YRDC deprivation and with or without threonine restriction (control media, 800 µM threonine; threonine-restricted media, 4 µM). To investigate the effects of YRDC and threonine purtubation on tRNA expression in GSCs, we performed tRNA sequencing in GSC456 with or without YRDC deprivation and with or without threonine restriction (control media, 800 µM threonine; threonine-restricted media, 4 µM). Overall design: Comparative gene expression profiling analysis of RNA-seq data, tRNA-seq data and comparative ribosome footprint analysis of ribosome profiling data for GSC456 with or without YRDC deprivation and with or without threonine restriction.