Quantitative effect of target translation on small RNA efficacy reveals a novel mode of interaction.
Bottom Line: Often the sRNA-binding site is adjacent to or overlapping with the ribosomal binding site (RBS), suggesting a possible interplay between sRNA and ribosome binding.Quantitative analysis of these data suggests a recruitment model, where bound ribosomes facilitate binding of the sRNA.Our findings offer a framework for understanding sRNA silencing in the context of bacterial physiology.
Affiliation: Department of Physics and FAS Center for Systems Biology, Harvard University, Cambridge, MA 02138, USA.Show MeSH
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Mentions: To characterize the relative ribosomal binding strength of the variants in our library we compared GFP expression without sRNA induction (0.5-mM IPTG, no aTc added) in a microplate reader. The different strains in our library demonstrated a wide range of GFP expression levels (Figure 3A, black bars), which correlated with their predicted ribosome binding strength (Supplementary Figure S9A). To measure the effect of the ribosome binding strength on sRNA efficacy, we repeated our measurements with induction of RyhB (IPTG = 0.5 mM, aTc = 8 ng/ml) (Figure 3A, red bars). In most strains, the presence of RyhB reduced GFP expression, as expected from the repression of sodB by RyhB.
Affiliation: Department of Physics and FAS Center for Systems Biology, Harvard University, Cambridge, MA 02138, USA.