Methylation Protects miRNAs and siRNAs from a 3′-End Uridylation Activity in Arabidopsis

Li, Junjie; Yang, Zhiyong; Yu, Bin; Li, Jun; Chen, Xuemei

doi:10.1016/j.cub.2005.07.029

Cited by 707 publications

(589 citation statements)

References 25 publications

(39 reference statements)

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“…In addition to the endogenous siRNAs, viruses and transgenes also give rise to siRNAs (reviewed in Brodersen and Voinnet, 2006;Vaucheret, 2006). We and others have shown that all types of siRNAs (except nat-siRNAs that have not been tested) are methylated in vivo in a HEN1-dependent manner (Akbergenov et al, 2006;Ebhardt et al, 2005;Li et al, 2005). Our biochemical studies with recombinant HEN1 protein also demonstrated that HEN1 is a siRNA methyltransferase (Yang et al, 2006).…”

Section: Introductionsupporting

confidence: 53%

Approaches for Studying MicroRNA and Small Interfering RNA Methylation In Vitro and In Vivo

Yang¹,

Vilkaitis²,

Yu³

et al. 2007

Methods in Enzymology

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The biogenesis of microRNAs (miRNAs) in plants is similar to that in animals, however, the processing of plant miRNAs consists of an additional step, the methylation of the miRNAs on the 3′ terminal nucleotides. The enzyme that methylates Arabidopsis miRNAs is encoded by a gene named HEN1, which has been shown genetically to be required for miRNA biogenesis in vivo. Small interfering RNAs (siRNAs) are also methylated in vivo in a HEN1-dependent manner. Our biochemical studies demonstrated that HEN1 is a methyltransferase acting on both miRNAs and siRNAs in vitro. HEN1 recognizes 21 to 24 nt small RNA duplexes, which are the products of Dicer-like enzymes, and transfers a methyl group from S-adenosylmethionine (SAM) to the 2′ OH of the last nucleotides of the small RNA duplexes. Here we describe methods to characterize the biochemical activities of the HEN1 protein both in vitro and in vivo, and methods to analyze the methylation status of small RNAs in vivo.

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Section: Introductionsupporting

confidence: 53%

Approaches for Studying MicroRNA and Small Interfering RNA Methylation In Vitro and In Vivo

Yang¹,

Vilkaitis²,

Yu³

et al. 2007

Methods in Enzymology

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“…Similarly, miR-16, a mammalian miRNA used as a control, was present primarily in the npg www.cell-research.com | Cell Research EVs. Plant miRNAs bear a single 2'-O-methylation on their 3′ end [23], which is necessary for their stability [38]. Since mammalian miRNAs lack this modification, we used sodium periodate oxidation to confirm that the miR159 in human serum is of plant origin.…”

Section: Plant Mir159 Can Be Detected In Human Sera and Tumor Tissuesmentioning

confidence: 99%

Cross-kingdom inhibition of breast cancer growth by plant miR159

Fong²,

et al. 2016

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MicroRNAs (miRNAs) are critical regulators of gene expression, and exert extensive impacts on development, physiology, and disease of eukaryotes. A high degree of parallelism is found in the molecular basis of miRNA biogenesis and action in plants and animals. Recent studies interestingly suggest a potential cross-kingdom action of plant-derived miRNAs, through dietary intake, in regulating mammalian gene expression. Although the source and scope of plant miRNAs detected in mammalian specimens remain controversial, these initial studies inspired us to determine whether plant miRNAs can be detected in Western human sera and whether these plant miRNAs are able to influence gene expression and cellular processes related to human diseases such as cancer. Here we found that Western donor sera contained the plant miRNA miR159, whose abundance in the serum was inversely correlated with breast cancer incidence and progression in patients. In human sera, miR159 was predominantly detected in the extracellular vesicles, and was resistant to sodium periodate oxidation suggesting the plant-originated 2'-O-methylation on the 3′ terminal ribose. In breast cancer cells but not non-cancerous mammary epithelial cells, a synthetic mimic of miR159 was capable of inhibiting proliferation by targeting TCF7 that encodes a Wnt signaling transcription factor, leading to a decrease in MYC protein levels. Oral administration of miR159 mimic significantly suppressed the growth of xenograft breast tumors in mice. These results demonstrate for the first time that a plant miRNA can inhibit cancer growth in mammals.

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“…Analogous biochemical work on the mouse HEN1 protein has shown that its activity is specific for single-stranded piRNAs (Kirino and Mourelatos 2007a), consistent with the lack of a dsRBD in the animal orthologs identified to date. Therefore, HEN1 proteins function to add methyl groups to the The blot on the left shows the laddering effect observed for miR167 in an Arabidopsis hen1-1 mutant versus a Landsberg erecta (Ler) control; reprinted from Li et al (2005). At top right, in wild-type (WT) plants, miRNAs are 3 0 methylated by HEN1, but may be decayed by the 3 0 to 5 0 exonuclease SDN1/2.…”

Section: Discovery Of Hen1 and Its Conserved Function In Many Speciesmentioning

confidence: 99%

Deep Sequencing from hen1 Mutants to Identify Small RNA 3' Modifications

Zhai

Meyers

2012

Cold Spring Harbor Symposia on Quantitative Biology

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microRNAs (miRNAs) function via targeting of messenger RNAs, suppressing protein levels, and playing important roles in biological processes of plants and animals. The pathway for miRNA biogenesis is well established, but less is known about miRNA turnover, largely because of difficulties in capturing miRNAs during the process of decay, in which they are both rare and ephemeral. The HEN1 protein methylates the 3 0 terminus of small RNAs (sRNAs), protecting them from poly-urydilation and degradation. Recent progress using deep sequencing to study sRNAs in hen1 reveals the potential for hen1 mutants to serve as a platform for studies of miRNA turnover, with the sequencing data providing unprecedented precision and detail in the characterization of 3 0 modifications.

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Methylation Protects miRNAs and siRNAs from a 3′-End Uridylation Activity in Arabidopsis

Cited by 707 publications

References 25 publications

Approaches for Studying MicroRNA and Small Interfering RNA Methylation In Vitro and In Vivo

Approaches for Studying MicroRNA and Small Interfering RNA Methylation In Vitro and In Vivo

Cross-kingdom inhibition of breast cancer growth by plant miR159

Deep Sequencing from hen1 Mutants to Identify Small RNA 3' Modifications

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