Three-parametric polysome profiling of Arabidopsis thaliana

In this work we extended the common method of polysome profiling by the additional measurement of the total mRNA fraction. Such three-parametric (polysome mRNA fraction, monosome mRNA fraction and total mRNA) profiling allowed to directly measure relative ribosomal load and directly normalize it to different total mRNA levels. Using customized bioinformatic analysis of the two-dimensional data of transcriptional and translational gene activities allowed to (i) deconvolute transcriptional and translational regulation; (ii) identify genes different by the level of translation independently from the level of transcription; (iii) quantify the influence of mRNA sequence characteristics on translational efficacy and (iv) find mRNA motifs in 5`UTR, specific to transcripts with high translation.