GSE98333
Analysis of Ribosome Protected Fragments of mRNAs from yeast strains either expressing Rio1p or depleted of Rio1p
mRNAs bound by ribosomes from yeast cells were analysed in order to determine the exact position of ribosomes in the presence or absence of Rio1p. Beside total Ribosome Protected Fragments (RPFs), RPFs from mRNAs protected by immature pre-40S pre-ribosomes was also analysed. The analysis showed that immature 40S ribosomes can carry out translation and their premature entry into translation is hindered by Rio1p. Overall design: mRNAs fragments protected by ribosomes were subjected to NGS Ribosome Profiling experiment. Samples included (1) total ribosome-associated mRNAs from yeast cells expressing Rio1p, (2) total ribosome-associated mRNAs from yeast cells depleted of Rio1p and (3) immuno-selected mRNAs associated with immature pre-40S pre-ribosomal particles from yeast-cells depleted of Rio1p. Ribosome-protected mRNA fragments were used as templates for cDNA libraries. The resulting cDNAs were sequenced and the sequences were annealed to the yeast genome. The position of the reads were determined in relation to either the translation start site or the stop codon of the corresponding coding DNA sequences.