Fig. 7
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- ZDB-FIG-210223-21
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- Tietz et al., 2020 - Pumilio response and AU-rich elements drive rapid decay of Pnrc2-regulated cyclic gene transcripts
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Fig. 7. The Pumilio response and AU-rich elements in the her1 3′UTR are both required for rapid reporter transcript turnover. (A–C) Transgenic embryos carrying the hsp70l:Venus-her1 3′UTR with disrupted PRE & ARE-SV40 pA reporter (line oz93) with a 2 nt mutation in the PRE sequence and 3 nt mutation in the ARE sequence were raised to mid-segmentation stage and heat shocked for 15 min, then collected at the indicated minutes pHS and processed by Venus in situ hybridization (n ≥ 11 embryos per time point). Venus transcript is not detected in the absence of heat shock (n = 10 embryos) (data not shown). (D) qPCR analysis comparing Venus transcript fold change from 30 min pHS to 60 and 90 min pHS for the reporter lines Tg(hsp70l:Venus-her1 3′UTR-SV40 pA)oz44 and Tg(hsp70l:Venus-her1 3′UTR with disrupted PRE & ARE-SV40 pA)oz93 (n = 10 embryos per time point across three biological replicates). The presence of both mutations extends reporter half-life by ~7-fold compared to the unmutated control. Three independent lines carrying the hsp70l:Venus-her1 3′UTR with disrupted PRE & ARE-SV40 pA reporter were analyzed by in situ hybridization and qPCR and each line exhibited comparable Venus decay dynamics (data not shown and Supplemental Fig. 5); one representative line is shown (see Methods for details). pHS = post-heat shock; PRE = Pumilio response element; ARE = AU-rich element; t1/2 = half-life; ± = standard deviation; pA = polyadenylation sequence. |
Reprinted from Developmental Biology, 462, Tietz, K.T., Gallagher, T.L., Mannings, M.C., Morrow, Z.T., Derr, N.L., Amacher, S.L., Pumilio response and AU-rich elements drive rapid decay of Pnrc2-regulated cyclic gene transcripts, 129-140, Copyright (2020) with permission from Elsevier. Full text @ Dev. Biol.