Fig. 5: Zygotic expression of miR-430 inhibits translation of mRNAs with 5’ UTR seed sites. | Nature Communications

Fig. 5: Zygotic expression of miR-430 inhibits translation of mRNAs with 5’ UTR seed sites.

From: NaP-TRAP reveals the regulatory grammar in 5’UTR-mediated translation regulation during zebrafish development

Fig. 5

a Cumulative distributions of translation (6 hpf / 2 hpf) for reporters containing miR-430 (GCACUUA, GCACUUU, AGCACUU; N = 212 reporters) or miR-1 (ACAUUCC, CAUUCCA; N = 71 reporters) heptamers, labeled in green and blue respectively. Reporters labeled in gray contain neither seed sequence (N = 8,242 reporters) (two-sided Mann-Whitney U test, p < 10−27 miR-430 reporters vs reporters with no seed; two-sided Mann-Whitney U test, p < 0.001 miR-1 reporters vs reporters with no seed). b, c The effect of the number of complementary bases to miR-430 ((b), N = 282 reporters) or miR-1 ((c), N = 185 reporters) on the translation of reporters containing seed sequences at 6 hpf versus 2 hpf (two-sided Pearson’s R; (b) p < 10−3, (c) p < 0.98). d–e The effect of the number of complementary bases to miR-430 ((d), N = 792 reporters) and miR-1 ((e), N = 425 reporters) on translation for reporters containing seeds with a single mismatch at 6 hpf versus 2 hpf (two-sided Pearson’s R; (d) p < 0.8, (e) p < 0.8). f, g Plots showing the delta translation (Translation 6 hpf / 2hpf) of reporter fragments that tile the zebrafish 5’ UTR (ENSDART00000135384 / usp9 (f), ENSDART00000063359 / ucp2 (g)). The seed sites of miR-430 (AGCAUUU) are labeled in green. h, i Schematic detailing dual luciferase assay measuring the inhibitory effect of miR-430 binding sites in the 5’ UTR (h). 4xmiR-430-nanoluc and 4xmiR-430-shuffled-nanoluc were injected in wild-type and miR-430 knockout embryos. Relative Luciferase Activity (RLU) values were normalized to each reporter (N = 3 replicates; 5 embryos per replicate; two-sided unpaired t-test; (i) p < 0.001; error bars = SD).

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