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Sequestration of microRNA-mediated target repression by the Ago2-associated RNA-binding protein FAM120A

Author(s)
Kelly, Timothy J.; Suzuki, Hiroshi I.; Zamudio, Jesse R.; Suzuki, Megumu
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Creative Commons Attribution NonCommercial License 4.0 https://creativecommons.org/licenses/by-nc/4.0/
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Abstract
Argonaute (Ago) proteins interact with various binding partners and play a pivotal role in microRNA (miRNA)-mediated silencing pathways. By utilizing immunoprecipitation followed by mass spectrometry to determine cytoplasmic Ago2 protein complexes in mouse embryonic stem cells (mESCs), we identified a putative RNA-binding protein FAM120A (also known as OSSA/C9ORF10) as an Ago2 interacting protein. Individual nucleotide resolution cross-linking and immunoprecipitation (iCLIP) analysis revealed that FAM120A binds to homopolymeric tracts in 3′′-UTRs of about 2000 mRNAs, particularly poly(G) sequences. Comparison of FAM120A iCLIP and Ago2 iCLIP reveals that greater than one-third of mRNAs bound by Ago2 in mESCs are co-bound by FAM120A. Furthermore, such FAM120A-bound Ago2 target genes are not subject to Ago2-mediated target degradation. Reporter assays suggest that the 3′′-UTRs of several FAM120A-bound miRNA target genes are less sensitive to Ago2-mediated target repression than those of FAM120A-unbound miRNA targets and FAM120A modulates them via its G-rich target sites. These findings suggest that Ago2 may exist in multiple protein complexes with varying degrees of functionality.
Date issued
2019-07-09
URI
https://hdl.handle.net/1721.1/124507
Department
Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Biology; Koch Institute for Integrative Cancer Research at MIT
Journal
RNA
Publisher
Cold Spring Harbor Laboratory Press
Citation
Kelly, Timothy J. et al. "Sequestration of microRNA-mediated target repression by the Ago2-associated RNA-binding protein FAM120A." RNA 25 (2019):1291-1297 © 2019 The Author(s)
Version: Final published version
ISSN
1355-8382
1469-9001
Keywords
Molecular Biology

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