IDEAS home Printed from https://ideas.repec.org/a/nat/nature/v542y2017i7640d10.1038_nature21025.html
   My bibliography  Save this article

An Argonaute phosphorylation cycle promotes microRNA-mediated silencing

Author

Listed:
  • Ryan J. Golden

    (University of Texas Southwestern Medical Center
    Medical Scientist Training Program, University of Texas Southwestern Medical Center)

  • Beibei Chen

    (Quantitative Biomedical Research Center, University of Texas Southwestern Medical Center
    University of Texas Southwestern Medical Center)

  • Tuo Li

    (University of Texas Southwestern Medical Center)

  • Juliane Braun

    (University of Texas Southwestern Medical Center)

  • Hema Manjunath

    (University of Texas Southwestern Medical Center)

  • Xiang Chen

    (University of Texas Southwestern Medical Center)

  • Jiaxi Wu

    (University of California San Francisco)

  • Vanessa Schmid

    (Eugene McDermott Center for Human Growth & Development, University of Texas Southwestern Medical Center)

  • Tsung-Cheng Chang

    (University of Texas Southwestern Medical Center)

  • Florian Kopp

    (University of Texas Southwestern Medical Center)

  • Andres Ramirez-Martinez

    (University of Texas Southwestern Medical Center)

  • Vincent S. Tagliabracci

    (University of Texas Southwestern Medical Center)

  • Zhijian J. Chen

    (University of Texas Southwestern Medical Center
    Howard Hughes Medical Institute, University of Texas Southwestern Medical Center)

  • Yang Xie

    (Quantitative Biomedical Research Center, University of Texas Southwestern Medical Center
    University of Texas Southwestern Medical Center
    Harold C. Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center)

  • Joshua T. Mendell

    (University of Texas Southwestern Medical Center
    Howard Hughes Medical Institute, University of Texas Southwestern Medical Center
    Harold C. Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center
    Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center)

Abstract

MicroRNAs (miRNAs) perform critical functions in normal physiology and disease by associating with Argonaute proteins and downregulating partially complementary messenger RNAs (mRNAs). Here we use clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) genome-wide loss-of-function screening coupled with a fluorescent reporter of miRNA activity in human cells to identify new regulators of the miRNA pathway. By using iterative rounds of screening, we reveal a novel mechanism whereby target engagement by Argonaute 2 (AGO2) triggers its hierarchical, multi-site phosphorylation by CSNK1A1 on a set of highly conserved residues (S824–S834), followed by rapid dephosphorylation by the ANKRD52–PPP6C phosphatase complex. Although genetic and biochemical studies demonstrate that AGO2 phosphorylation on these residues inhibits target mRNA binding, inactivation of this phosphorylation cycle globally impairs miRNA-mediated silencing. Analysis of the transcriptome-wide binding profile of non-phosphorylatable AGO2 reveals a pronounced expansion of the target repertoire bound at steady-state, effectively reducing the active pool of AGO2 on a per-target basis. These findings support a model in which an AGO2 phosphorylation cycle stimulated by target engagement regulates miRNA:target interactions to maintain the global efficiency of miRNA-mediated silencing.

Suggested Citation

  • Ryan J. Golden & Beibei Chen & Tuo Li & Juliane Braun & Hema Manjunath & Xiang Chen & Jiaxi Wu & Vanessa Schmid & Tsung-Cheng Chang & Florian Kopp & Andres Ramirez-Martinez & Vincent S. Tagliabracci &, 2017. "An Argonaute phosphorylation cycle promotes microRNA-mediated silencing," Nature, Nature, vol. 542(7640), pages 197-202, February.
  • Handle: RePEc:nat:nature:v:542:y:2017:i:7640:d:10.1038_nature21025
    DOI: 10.1038/nature21025
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/nature21025
    File Function: Abstract
    Download Restriction: Access to the full text of the articles in this series is restricted.

    File URL: https://libkey.io/10.1038/nature21025?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    As the access to this document is restricted, you may want to search for a different version of it.

    Citations

    Citations are extracted by the CitEc Project, subscribe to its RSS feed for this item.
    as


    Cited by:

    1. Zhangli Su & Ida Monshaugen & Briana Wilson & Fengbin Wang & Arne Klungland & Rune Ougland & Anindya Dutta, 2022. "TRMT6/61A-dependent base methylation of tRNA-derived fragments regulates gene-silencing activity and the unfolded protein response in bladder cancer," Nature Communications, Nature, vol. 13(1), pages 1-17, December.
    2. Tingting Niu & Charlotte Rosny & Séverine Chautard & Amaury Rey & Danish Patoli & Marine Groslambert & Camille Cosson & Brice Lagrange & Zhirong Zhang & Orane Visvikis & Sabine Hacot & Maggy Hologne &, 2021. "NLRP3 phosphorylation in its LRR domain critically regulates inflammasome assembly," Nature Communications, Nature, vol. 12(1), pages 1-15, December.
    3. Tian-Yu Song & Min Long & Hai-Xin Zhao & Miao-Wen Zou & Hong-Jie Fan & Yang Liu & Chen-Lu Geng & Min-Fang Song & Yu-Feng Liu & Jun-Yi Chen & Yu-Lin Yang & Wen-Rong Zhou & Da-Wei Huang & Bo Peng & Zhen, 2021. "Tumor evolution selectively inactivates the core microRNA machinery for immune evasion," Nature Communications, Nature, vol. 12(1), pages 1-15, December.
    4. Charlotte A. Cialek & Gabriel Galindo & Tatsuya Morisaki & Ning Zhao & Taiowa A. Montgomery & Timothy J. Stasevich, 2022. "Imaging translational control by Argonaute with single-molecule resolution in live cells," Nature Communications, Nature, vol. 13(1), pages 1-14, December.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:nature:v:542:y:2017:i:7640:d:10.1038_nature21025. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.