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| Status |
Public on May 17, 2024 |
| Title |
Epigenetic regulation by dynamic RNA G-quadruplex folding and unfolding (DMS-Seq) |
| Organism |
Mus musculus |
| Experiment type |
Other
|
| Summary |
Whether RNA G-quadruplexes (rG4) form extensively in vivo and what roles they play remain actively debated. Among their proposed functions is recognition of Polycomb repressive complex 2 (PRC2), but how the interaction results in epigenetic regulation is not understood. Here we demonstrate that rG4s form dynamically during ES cell differentiation and require ATRX’s helicase function to unwind competing secondary structures. Mutating ATRX causes rG4 depletion on a transcriptome-wide basis and dramatically increases gene expression. We identify and mutate rG4s within Xist RNA and mechanistically separate PRC2’s recruitment versus catalysis. Surprisingly, although rG4s recruit PRC2, unfolding the rG4 structure causes PRC2 hyperactivation, entrapment of PRC2 in the S1 chromosomal compartment, and loss of gene silencing. Thus, we link dynamic rG4 folding and unfolding to PRC2 recruitment, trans-compartmental Xist migration, regulated activation of PRC2, and whole-chromosome gene silencing.
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| Overall design |
Modified existing rG4-seq and RT-Stop protocols were performed and generated an “in vitro” and “in vivo” pipeline, based on differential sensitivity of various RNA structures to dimethyl sulfate (DMS), reverse transcriptase (RT), and the monovalent cations Li+ versus K+.
|
| Web link |
https://doi.org/10.1016/j.molcel.2024.04.015
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| |
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| Contributor(s) |
Lee YW, Weissbein U, Blum R, Lee JT |
| Citation(s) |
38759625 |
| |
| Submission date |
Nov 30, 2022 |
| Last update date |
Aug 16, 2024 |
| Contact name |
yongwoo lee |
| E-mail(s) |
ylee@molbio.mgh.harvard.edu
|
| Phone |
6177552856
|
| Organization name |
MASSACHUSETTS GENERAL HOSPITAL
|
| Department |
Mol bio
|
| Lab |
Jeannie Lee Lab
|
| Street address |
185 Cambridge St
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02114 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
|
| Samples (6)
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| This SubSeries is part of SuperSeries: |
| GSE219083 |
G-quadruplex folding in Xist RNA antagonizes PRC2 activity for step-wise regulation of X-chromosome inactivation |
|
| Relations |
| BioProject |
PRJNA907004 |
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