Argonaute-miRNA Complexes Silence Target mRNAs in the Nucleus of Mammalian Stem Cells

Sarshad, Aishe A.; Juan, Aster H.; Muler, Ana Iris Correa; Anastasakis, Dimitrios G.; Wang, Xiantao; Genzor, Pavol; Feng, Xuesong; Tsai, Pei-Fang; Sun, Hongwei; Haase, Astrid D.; Sartorelli, Vittorio; Hafner, Markus

doi:10.1016/j.molcel.2018.07.020

Cited by 114 publications

(148 citation statements)

References 88 publications

(140 reference statements)

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“…Previous studies have demonstrated that 3'-untranslated regions (3'UTRs) of mRNAs are the preferred, although not exclusive, sites of interaction between miRNAs and mRNAs (Bartel, 2018;Chi et al, 2009;Sarshad et al, 2018). Consistent with these findings, the majority of HEAP peaks we identified in mESCs mapped to 3'UTRs, followed by sites mapping to coding exons (Figure 2a, Supplementary Figure 2c).…”

Section: A Halo-ago2 Fusion Protein Enables Antibody-free Purificatiosupporting

confidence: 87%

High-resolution in vivo identification of miRNA targets by Halo-Enhanced Ago2 Pulldown

Pritykin

Concepcion

et al. 2019

Preprint

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The identification of miRNA targets by Ago2 crosslinking-immunoprecipitation (CLIP) methods has provided major insights into the biology of this important class of non-coding RNAs. However, these methods are technically challenging and not easily translated to an in vivo setting. To overcome these limitations and to facilitate the investigation of miRNA functions in mice, we have developed a method (HEAP: for Halo-Enhanced Ago2 Pulldown) to map miRNA-mRNA binding sites. This method is based on a novel genetically engineered mouse harboring a conditional, Cre-regulated, Halo-Ago2 allele expressed from the endogenous Ago2 locus. By using a resin conjugated to the HaloTag ligand, Ago2-miRNA-mRNA complexes can be efficiently purified from cells and tissues expressing the endogenous Halo-Ago2 allele. We demonstrate the reproducibility and sensitivity of this method in mouse embryonic stem cells, in developing embryos, in adult tissues and in autochthonous mouse models of human brain and lung cancers. The tools and the datasets we have generated will serve as a valuable resource to the scientific community and will facilitate the characterization of miRNA functions under physiological and pathological conditions.

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Section: A Halo-ago2 Fusion Protein Enables Antibody-free Purificatiosupporting

confidence: 87%

High-resolution in vivo identification of miRNA targets by Halo-Enhanced Ago2 Pulldown

Pritykin

Concepcion

et al. 2019

Preprint

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“…Human genome encodes four Argonaute proteins, Ago1-4. A recent report shows that Argonaute-miRNA complexes can target mRNAs even in the nucleus of stem cells (Sarshad et al, 2018). Ago proteins interact with miRNAs and siRNAs and load them onto their target mRNAs to silence their expression.…”

Section: Introductionmentioning

confidence: 99%

“…They are highly conserved proteins and found in most eukaryotes (except Saccharomyces cerevisiae). A recent report shows that Argonaute-miRNA complexes can target mRNAs even in the nucleus of stem cells (Sarshad et al, 2018). In addition to their role in small RNA-mediated post-transcriptional gene silencing, human Ago proteins have roles in transcription, splicing, and DNA repair (Meister, 2013).…”

Section: Introductionmentioning

confidence: 99%

Let‐7a‐regulated translational readthrough of mammalian AGO 1 generates a micro RNA pathway inhibitor

et al. 2019

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Translational readthrough generates proteins with extended C‐termini, which often possess distinct properties. Here, we have used various reporter assays to demonstrate translational readthrough of AGO1 mRNA. Analysis of ribosome profiling data and mass spectrometry data provided additional evidence for translational readthrough of AGO1. The endogenous readthrough product, Ago1x, could be detected by a specific antibody both in vitro and in vivo. This readthrough process is directed by a cis sequence downstream of the canonical AGO1 stop codon, which is sufficient to drive readthrough even in a heterologous context. This cis sequence has a let‐7a miRNA‐binding site, and readthrough is promoted by let‐7a miRNA. Interestingly, Ago1x can load miRNAs on target mRNAs without causing post‐transcriptional gene silencing, due to its inability to interact with GW182. Because of these properties, Ago1x can serve as a competitive inhibitor of miRNA pathway. In support of this, we observed increased global translation in cells overexpressing Ago1x. Overall, our results reveal a negative feedback loop in the miRNA pathway mediated by the translational readthrough product of AGO1.

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“…However, there is accumulating evidence that some fraction of mammalian miRNAs may also execute important nuclear regulatory functions by controlling gene expression in the nucleus at both transcriptional and posttranscriptional levels, as well as by affecting alternative splice site selection (Roberts, 2014). In a set of elegant experiments, a significant fraction of miRNAs and several RISC components, including AGOs, have been identified to become located and functionally operative within the nucleus of human cells (Avivi et al, 2017; Bottini et al, 2017; Castanotto et al, 2018; Gagnon, Li, Chu, Janowski, & Corey, 2014; Ohrt et al, 2008; Sarshad et al, 2018; Weinmann et al, 2009). Nuclear localized miRNAs were found capable both to suppress and to stimulate transcriptional expression at distinct gene loci, involving direct and indirect interference pathways.…”

Section: Epigenetic Foundations Of Transcriptional Memorymentioning

confidence: 99%

Transcriptional memory in skeletal muscle. Don't forget (to) exercise

Beiter

Nieß

Moser

2020

Journal Cellular Physiology

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Transcriptional memory describes an ancient and highly conserved form of cellular learning that enables cells to benefit from recent experience by retaining a mitotically inheritable but reversible memory of the initial transcriptional response when encountering an environmental or physiological stimulus. Herein, we will review recent progress made in the understanding of how cells can make use of diverse constituents of the epigenetic toolbox to retain a transcriptional memory of past states and perturbations. Specifically, we will outline how these mechanisms will help to improve our understanding of skeletal muscle plasticity in health and disease. We describe the epigenetic road map that allows skeletal muscle fibers to navigate through training‐induced adaptation processes, and how epigenetic memory marks can preserve an autobiographical history of lifestyle behavior changes, pathological challenges, and aging. We will further consider some key findings in the field of exercise epigenomics to emphasize major challenges when interpreting dynamic changes in the chromatin landscape in response to acute exercise and training.

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Argonaute-miRNA Complexes Silence Target mRNAs in the Nucleus of Mammalian Stem Cells

Cited by 114 publications

References 88 publications

High-resolution in vivo identification of miRNA targets by Halo-Enhanced Ago2 Pulldown

High-resolution in vivo identification of miRNA targets by Halo-Enhanced Ago2 Pulldown

Let‐7a‐regulated translational readthrough of mammalian AGO 1 generates a micro RNA pathway inhibitor

Transcriptional memory in skeletal muscle. Don't forget (to) exercise

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