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Updated: Jun 7, 2025
Visualization of G3BP Stress Granules Dynamics in Live Primary Cells
Published on: May 21, 2014
Decoding the interplay between m6A modification and stress granule stability by live-cell imaging
Qianqian Li1, Jian Liu1, Liping Guo1,2
1Shenzhen Bay Laboratory, Shenzhen 518132, China.
Abstract:
N6-methyladenosine (m6A)-modified mRNAs and their cytoplasmic reader YTHDFs are colocalized with stress granules (SGs) under stress conditions, but the interplay between m6A modification and SG stability remains unclear. Here, we presented a spatiotemporal m6A imaging system (SMIS) that can monitor the m6A modification and the translation of mRNAs with high specificity and sensitivity in a single live cell. SMIS showed that m6A-modified reporter mRNAs dynamically enriched into SGs under arsenite stress and gradually partitioned into the cytosol as SG disassembled. SMIS revealed that knockdown of YTHDF2 contributed to SG disassembly, resulting in the fast redistribution of mRNAs from SGs and rapid recovery of stalled translation. The mechanism is that YTHDF2 can regulate SG stability through the interaction with G3BP1 in m6A-modified RNA-dependent manner. Our results suggest a mechanism for the interplay between m6A modification and SG through YTHDF2 regulation.
Insights
N6-methyladenosine (m6A) modification and its reader YTHDF2 regulate stress granule (SG) stability. YTHDF2
Area of Science:
- Molecular Biology
- Cell Biology
- RNA Biology
Background:
- N6-methyladenosine (m6A) modification and its cytoplasmic reader proteins, YTHDFs, are known to interact with stress granules (SGs).
- The precise role of m6A modification and YTHDF proteins in regulating SG dynamics and mRNA translation under stress remains largely uncharacterized.
Purpose of the Study:
- To investigate the interplay between m6A modification and stress granule stability.
- To elucidate the mechanism by which YTHDF proteins influence SG dynamics and mRNA translation recovery.
Main Methods:
- Development and application of a spatiotemporal m6A imaging system (SMIS) for live-cell monitoring of m6A modification and mRNA translation.
- Utilizing SMIS to observe dynamic changes in m6A-modified mRNAs within SGs under arsenite stress.
- Employing knockdown of YTHDF2 to assess its impact on SG disassembly and mRNA redistribution.
Main Results:
- SMIS demonstrated dynamic enrichment of m6A-modified mRNAs into SGs under arsenite stress, followed by partitioning into the cytosol upon SG disassembly.
- Knockdown of YTHDF2 accelerated SG disassembly, leading to faster mRNA redistribution and recovery of stalled translation.
- YTHDF2 was found to regulate SG stability via interaction with G3BP1 in an m6A-modified RNA-dependent manner.
Conclusions:
- m6A modification and YTHDF2 play a crucial role in regulating stress granule stability.
- YTHDF2's interaction with G3BP1, dependent on m6A-modified RNA, mediates SG stability and translation recovery.
- This study reveals a novel mechanism linking m6A modification to stress granule dynamics and translational control.
