Ren Guodong, Wang Xiaoyan, Yu Bin
State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai, 200438, China.
Center for Plant Science Innovation and School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, 68588-0660, NE, USA.
Methods Mol Biol. 2017;1640:23-37. doi: 10.1007/978-1-4939-7165-7_2.
Uridylation (3' untemplated uridine addition) provides a mechanism to trigger the degradation of miRNAs and the 5' cleavage products (5' CP) that are produced from miRNA-directed ARGONAUTE (AGO) cleavage of target RNAs. We have recently shown that HEN1 SUPPRESSOR 1 (HESO1), a terminal uridylyltransferase, and its homolog UTP:RNA uridylyltransferase 1 (URT1) catalyze the uridylation of miRNAs and 5' CPs within the AGO complex in higher plants. In this chapter, we describe detailed protocols for analyzing 3' end uridylation of both AGO-bound miRNAs and 5' CP.
尿苷化（3'非模板尿苷添加）提供了一种机制，可触发微小RNA（miRNA）以及由miRNA指导的AGO（AGO）对靶RNA切割产生的5'切割产物（5'CP）的降解。我们最近表明，HEN1抑制因子1（HESO1），一种末端尿苷酰转移酶，及其同系物UTP：RNA尿苷酰转移酶1（URT1），在高等植物的AGO复合物中催化miRNA和5'CP的尿苷化。在本章中，我们描述了分析AGO结合的miRNA和5'CP的3'末端尿苷化的详细方案。
