TUT4/7-dependent miRNAs gain adenylation upon loss of uridylation. (A) The 133 miRNAs out of the 167 overlapping modified miRNAs in DU145 and IGROV1 are depicted, and each miRNA has a terminal modification frequency of at least >1%. Fold change is calculated in the TUT4/7 double mutants compared to the wild-type control. The x-axis denotes logarithmic (base 2) fold-change in uridylation while the y-axis displays logarithmic (base 2) fold-change in adenylation in cKO versus WT. The miRNAs specific to IGROV1 are depicted in yellow and miRNAs specific to DU145 are displayed in blue. The labeled miRNAs are the ones with at least fivefold increase in adenylation. (B) The y-axis indicates mean isomiR percentage. Both let-7b and let-7i have a higher 5p mature miRNA abundance compared to the 3p mature miRNA abundance, that is, a 5p/3p ratio of >1. Proportion of canonical, adenylated, and uridylated isomiRs of let-7b-3p and let-7i-3p is displayed. (C) Proportion of canonical, adenylated, and uridylated isomiRs of miR-652-3p and miR-103a-3p with 5p/3p < 1 is displayed. (D–G) Abundances of 3p mature miRNAs of let-7b-3p (D), let-7i-3p (E), miR-652-3p (F), and miR-103a-2 (G) in the WT controls of DU145 and IGROV1 with their corresponding TUT4/7 double mutants (catalytic knockouts) represented by cKOs.
