Skip to main content
. Author manuscript; available in PMC: 2016 Jul 16.
Published in final edited form as: Mol Cell. 2015 Jul 2;59(2):217–228. doi: 10.1016/j.molcel.2015.05.034

Figure 3.

Figure 3

Tailor is a uridyltransferase that inhibits mirtron biogenesis and activity. (A) Immunopurified wildtype and catalytically inactive myc-Tailor from S2 cells, detected with anti-myc, used for subsequent tailing assays. (B) Timecourse assay of myc-Tailor incubated with radiolabeled mature miR-1010 in the presence of uridine. Its robust tailing activity is dependent on its catalytic site. (C) Tailor only generates extensively tailed products using uridine; 5′ reaction time. (D) Tailor is dependent on Mg++ ions. (E) Tailor is highly efficient on a mirtron hairpin (pre-mir-1010) substrate. (F, G) Reporter assays in S2 cells transiently expressing ub-Gal4, UAS-mir-1003 (mirtron) and the indicated luciferase sensors. Tailor knockdown enhances mirtron activity (F) while Ectopic Tailor antagonizes mirtron activity (G). Error bars represent SD and unpaired T-tests were used to calculate significance. (H) In vitro dicing of unmodified and uridylated pre-mir-1010 hairpins by recombinant Dcr-1/Loqs-PB. Tailing inhibits (for 2U) or abolishes (for 4U) the accumulation of mature species. (I) Quantification of three dicing assays; standard deviations are shown. See also Figure S3.