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| Status |
Public on Jul 26, 2021 |
| Title |
FACT is recruited to the +1 nucleosome of transcribed genes and spreads in a Chd1-dependent manner [ChIP-exo] |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
The histone chaperone FACT occupies transcribed regions where it plays prominent roles in maintaining chromatin integrity and preserving epigenetic information. How it is targeted to transcribed regions, however, remains unclear. Proposed models for how FACT finds its way to transcriptionally active chromatin include docking on the RNA polymerase II (RNAPII) C-terminal domain (CTD), recruitment by elongation factors, recognition of modified histone tails and binding partially disassembled nucleosomes. Here, we systematically tested these and other scenarios in Saccharomyces cerevisiae and found that FACT binds transcribed chromatin, not RNAPII. Through a combination of experimental and mathematical modeling evidence, we propose that FACT recognizes the +1 nucleosome, as it is partially unwrapped by the engaging RNAPII, and spreads to downstream nucleosomes aided by the chromatin remodeler Chd1. Our work clarifies how FACT interacts with genes, suggests a processive mechanism for FACT function, and provides a framework to further dissect the molecular mechanisms of transcription-coupled histone chaperoning.
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| Overall design |
Paired-end sequencing of exonuclease-treated chromatin immunoprecipitation (ChIP-exo) of both FACT subunits (Spt16 and Pob3), RNAPII (Rpb3) and rabbit IgG in wild type and chd1Δ cells, from two independent biological replicates (except for the IgG ChIPs which were done once), for a total of 14 ChIP-exo samples.
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| Contributor(s) |
Jeronimo C, Angel A, Poitras C, Collin P, Mellor J, Robert F |
| Citation(s) |
34380014 |
| |
| Submission date |
Jul 27, 2020 |
| Last update date |
Oct 25, 2021 |
| Contact name |
Francois Robert |
| E-mail(s) |
francois.robert@ircm.qc.ca
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| Organization name |
IRCM
|
| Lab |
Chromatin and Genomic Expression
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| Street address |
110 av des Pins Ouest
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| City |
Montreal |
| State/province |
QC |
| ZIP/Postal code |
H2W 1R7 |
| Country |
Canada |
| |
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| Platforms (1) |
| GPL21656 |
Illumina HiSeq 4000 (Saccharomyces cerevisiae) |
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| Samples (14)
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| This SubSeries is part of SuperSeries: |
| GSE155144 |
FACT is recruited to the +1 nucleosome of transcribed genes and spreads in a Chd1-dependent manner |
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| Relations |
| BioProject |
PRJNA648838 |
| SRA |
SRP273674 |
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