URT1-mediated uridylation shapes poly(A) tails in Arabidopsis

The goal of this study was to evaluate the impact of URT1 on poly(A) tail profiles in Arabidopsis thaliana. Flowers were harvested from wild-type and urt1-1 (Salk_087647C) plants, three biological replicates each, grown on soil with 16 h light / 8 h darkness cycles (21/18°C). All Arabidopsis thaliana plants used in this study are of Columbia (Col-0) accession. Total RNA was extracted using Tri-Reagent (Molecular Research Center) from flowers, followed by mRNA enrichment with GST-eIF4E-K119A protein, according to the manufacturer's recommendations. Nanopore direct-RNA libraries were prepared from cap-enriched mRNA, mixed with poly(A)+ S. cerevisiae RNA, with Direct RNA Sequencing Kit (ONT, SQK-RNA002). Sequencing was performed with MinION device (Flow cell type R9.4.1; RevC) and basecalled using Guppy 4.0.11 (Oxford Nanopore Technologies).