Respective Contributions of URT1 and HESO1 to the Uridylation of 5' Fragments Produced From RISC-Cleaved mRNAs.

ABSTRACT: In plants, post-transcriptional gene silencing (PTGS) represses gene expression by translation inhibition and cleavage of target mRNAs. The slicing activity is provided by argonaute 1 (AGO1), and the cleavage site is determined by sequence complementarity between the target mRNA and the microRNA (miRNA) or short interfering RNA (siRNA) loaded onto AGO1, to form the core of the RNA induced silencing complex (RISC). Following cleavage, the resulting 5' fragment is modified at its 3' end by the untemplated addition of uridines. Uridylation is proposed to facilitate RISC recycling and the degradation of the RISC 5'-cleavage fragment. Here, we detail a 3' RACE-seq method to analyze the 3' ends of 5' fragments produced from RISC-cleaved transcripts. The protocol is based on the ligation of a pri

SUBMITTER: Zuber H 

PROVIDER: S-EPMC6191825 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature