Extended Data Fig. 4: Role of GHR in the production of 22-nt miRNAs.
From: Key RNA elements influencing DCL1 cleavage in plant microRNA biogenesis
(a) The plots display the average GHR motif scores in 3-bp motifs at various positions within pri-miR828 from diverse plants. (b) Diagrammatic structures of pri-miR828. The blue arrowheads indicate DCL1 cleavage sites. (c) The diagram indicates the lengths of potential fragments derived from pri-miR828 processing by BTL or LTB directions. (d) In vitro DCL1 cleavage assays were performed for pri-miR828 using two mutant DCL1 enzymes. The assays were repeated two times with similar results. (e) The repeated assay from Fig. 4d. (f) Time-course in vitro DCL1 cleavage assays were conducted for pre-miRNAs with different GHR scores. (g) Quantitative analysis of relative cleavage efficiency was derived from three repeated assays in (f), calculated from the band density of the cleaved product (F2 fragment) normalized to the high-GHR pre-miR828 at 3 min. The grey and green lines correspond to lowGHR, and highGHR of pre-miR828, respectively. Shaded regions around the lines represent 95% confidence intervals for cleavage efficiency measurements. (h) The DCL1 cleavage accuracy for WT or mutant pre-miR828 was calculated from the sequencing results of purified F1, F2, or F3 fragments obtained from the gel in Fig. 4d.
