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. Author manuscript; available in PMC: 2020 May 20.
Published in final edited form as: Dev Cell. 2019 Apr 4;49(4):542–555.e9. doi: 10.1016/j.devcel.2019.03.003

Figure 2. lep-5 is a long non-coding RNA.

Figure 2.

(A) The secondary structure of the lep-5 lncRNA as predicted by Turbofold. The conserved 5’ stem-loop (red bracket) includes the SL1 trans-spliced leader (in red). The 3’ stem-loop (blue bracket) and two base-paired regions forming the “central zipper” (green brackets) are also conserved. Dashed lines: boundaries of two hairpins deleted in the rescue experiments (Δ1 and Δ2). The polyA tail is omitted for clarity. For alignments, see Fig. S3 and STAR Methods. The 5’ region from predicted full-length structures of wild-type lep-5, lep-5(fs8 = G23A) and lep-5(G23A, C30T). Asterisks indicate position 23. SL1 is in red. See also Figures S4, S5. (C) Rescue experiments with transgenes containing stop codons in the predicted coding region (*3 or *2 stops), the lep-5 orthologs of C. briggsae and C. angaria and constructs containing nucleotide substitutions that discrupt and restore the lep-5 secondary structure (high lep-5(G23A) = transgene at 10x concentration). (D) Schematic of the SL1-spliced lep-5 RNA with putative ORFs shown above and selected predicted secondary structure features shown below. Boxes show the sequence of various mutant alleles compared with the wild-type allele. (E) Male tail tip phenotype at 20ºC of lep-5 mutants in which the secondary structure and/or the predicted ORFs are disrupted.