E-GEOD-48680 - Glucocorticoid effect on mRNA translation in childhood acute lymphoblastic leukemia
Released on 3 December 2013, last updated on 9 December 2013
Glucocorticoids (GCs) are a central component in treating childhood acute lymphoblastic leukemia (chALL). They mainly act via regulating gene transcription. However, control of mRNA translation by GC has never been assessed systematically. In our research, T- and precursor B-ALL cells were cultured with and without GC for 6 hours and subjected to translational profiling, a technique combining sucrose gradient fractionation and microarray analysis of mRNA in different fractions. Analysis of GC regulation in different pools revealed no significant differences in regulation of mRNA translation by GC, suggesting no evidence for translational regulation by GC. Ribosome profiling by sucrose gradient fractionation followed by microarray analysis of RNA from 3 distinct pools representing free RNA (pool 1), RNA in small protein-RNA complexes or bound to single ribosomes (pool 2) and RNA from large protein-RNA complexes (polysomes, pool 3). See Material and Methods of the original article for a detailed description. Translational efficiency of genes was estimated by comparing the amount of a gene's mRNA in pool 3 with the total amount of mRNA of the gene (in all 3 pools). Translational regulation by GC was estimated by contrasting gene regulations in pool 3 with gene regulation between GC-treated and control samples in all 3 pools (depicted as transcriptional regulation). Moderated t-tests were used to assess significance for translational regulation.
transcription profiling by array
Johannes Rainer <email@example.com>, Daniel Bindreither, Reinhard Kofler, Tatsiana Aneichyk
Translational profiling in childhood acute lymphoblastic leukemia: no evidence for glucocorticoid regulation of mRNA translation. Aneichyk T, Bindreither D, Mantinger C, Grazio D, Goetsch K, Kofler R, Rainer J. , Europe PMC 24289529