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Synergistic and independent actions of multiple terminal nucleotidyl transferases in 3’ tailing of small RNAs in Arabidopsis

ABSTRACT: Small silencing RNAs are key regulators of gene expression in both plants and animals. HEN1-mediated 3’ terminal 2’-O-methylation plays a crucial role in small RNA stability control. In the absence of HEN1, small RNAs are frequently uridylated (untemplated uridine addition) and trimmed, a phenomenon that is conserved across species. However, the underlying molecular mechanism is largely unknown. In this study, we have discovered UTP: RNA uridylyltransferase (URT1) acts redundantly with HESO1 in the uridylation of miRNAs, in addition to its role in oligo-adenylated mRNA uridylation. We show both common and distinctive features of URT1 and HESO1 in catalyzing miRNA uridylation.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE60826 | GEO | 2015/07/01

SECONDARY ACCESSION(S): PRJNA259631

REPOSITORIES: GEO

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Synergistic and independent actions of multiple terminal nucleotidyl transferases in 3’ tailing of small RNAs in Arabidopsis

Project description:Small silencing RNAs are key regulators of gene expression in both plants and animals. HEN1-mediated 3’ terminal 2’-O-methylation plays a crucial role in small RNA stability control. In the absence of HEN1, small RNAs are frequently uridylated (untemplated uridine addition) and trimmed, a phenomenon that is conserved across species. However, the underlying molecular mechanism is largely unknown. In this study, we have discovered UTP: RNA uridylyltransferase (URT1) acts redundantly with HESO1 in the uridylation of miRNAs, in addition to its role in oligo-adenylated mRNA uridylation. We show both common and distinctive features of URT1 and HESO1 in catalyzing miRNA uridylation. non coding RNA deep sequencing

2015-07-01 | E-GEOD-60826 | biostudies-arrayexpress

Distinct and cooperative activities of HESO1 and URT1 nucleotidyl transferases in microRNA turnover in Arabidopsis

Project description:3’ uridylation is increasingly recognized as a conserved RNA modification process associated with RNA turnover in eukaryotes. 2’-O-methylation on the 3’ terminal ribose protects micro(mi)RNAs from 3’ truncation and 3’ uridylation in Arabidopsis. Previously, we identified HESO1 as the nucleotidyl transferase that uridylates most unmethylated miRNAs in vivo, but substantial 3’ tailing of miRNAs still remains in heso1 loss-of-function mutants. In this study, we found that among nine HESO1 paralogs, UTP:RNA URIDYLYLTRANSFERASE 1 (URT1) is the single most predominant nucleotidyl transferase that tails miRNAs. URT1 and HESO1 prefer substrates with different 3’ end nucleotides in vitro and act cooperatively to tail different forms of the same miRNAs in vivo. Moreover, both HESO1 and URT1 exhibit nucleotidyl transferase activity on AGO1-bound miRNAs. Although these enzymes are able to add long tails to AGO1-bound miRNAs, the tailed miRNAs remain associated with AGO1. In fact, we show that a tailed miRNA acquires the ability to trigger the production of secondary siRNAs. Therefore, 3’ uridylation could endow new properties to miRNAs in addition to its known effects in miRNA degradation.

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NTP4 modulates miRNA accumulation via asymmetric modification of miRNA-3p/5p duplex

Project description:Arabidopsis uridylyltransferase on unmethylated sRNAs and directed their degradation. Here, the small RNA deep sequencing of the uridylyltransferase mutants of Arabidopsis were performed, which include Col-0, heso1-1, urt1-1, ntp4-1.

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Terminal nucleotidyl transferases govern secondary siRNA production at distinct steps in plants

Project description:In this study, we aim at understanding the contributions from TUTases, HESO1 and URT1, in miRNA-targeted secondary siRNA production in Arabidopsis thaliana. We have sequenced small RNAs from various T-DNA insertation single/double/triple mutants and compared their capability of producing ectopic secondary siRNAs from endogenous miRNA targets. The DESeq analysis is also included.

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High density analysis of the tailing and trimming patterns of RISC-cleaved MYB33 and SPL13 transcripts in Arabidopsis heso1 and urt1 mutants

Project description:Using 3' RACEseq method, we compared the tailing and nibbling patterns of RISC-cleaved MYB33 and SPL13 transcripts between wild-type plants and mutant plants depleted for the terminal uridylyltransferases (TUTases) HESO1 and URT1. Our data reveal a major and minor role for HESO and URT1, respectively, in the uridylation of RISC-cleaved transcripts.

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Nucleotidyltransferase gene HESO1

Project description:Small RNAs in plants are protected by 2'-O-methylation at 3' terminal nucleotide by HEN1. In hen1 mutant, small RNAs lacking 3' methylation are truncated and tailed at 3' end. We identified a nucleotidyltransferase gene HESO1 in Arabidopsis that tails small RNAs in hen1. In order to study the small RNA tailing and truncation in detail in wild type Col, heso1-1, hen1-8 and hen1-8 heso1-1 double mutant, we constructed small RNA libraries for these four genotypes and we found heso1-1 mutation reduces small RNA tailing in hen1-8 background, however, the 3' truncation is not significantly affected. 8 samples examined: wild-type Col (replicate 1), heso1-1 (replicate 1), hen1-8(replicate 1), hen1-8 heso1-1 (replicate 1), wild-type Col (replicate 2), heso1-1 (replicate 2), hen1-8(replicate 2) and hen1-8 heso1-1 (replicate 2).

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Impact of URT1-mediated uridylation on small RNA tailing in Arabidopsis

2016-03-11 | GSE72456 | GEO

Nucleotidyltransferase gene HESO1

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