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| Status |
Public on Sep 26, 2012 |
| Title |
Effects of Histone H3 depletion on nucleosome occupancy and positioning through the S. cerevisiae genome [Paired-end Mnase-seq] |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Experiments performed over the past three decades have shown that nucleosomes are transcriptional repressors. In Saccharomyces cerevisiae, depletion of histone H4 results in the genome-wide transcriptional de-repression of hundreds genes. The mechanism of de-repression is hypothesized to be rooted directly in chromatin changes. To test this, we reproduced classical H4 depletion experiments by conditional repression of all histone H3 transcription, which depletes the supply of nucleosomes in vivo. RNA-seq results were consistent with the earlier studies, but much more sensitive, revealing nearly 2500 de-repressed genes. Changes in chromatin organization were determined by MNase-seq. Nucleosomes that were preferentially retained occurred in regions of high DNA-encoded nucleosome affinity, and were marked with H3K36me2, which is linked to transcription elongation. Nucleosomes harboring acetyl marks or that contained the variant histone H2A.z were preferentially lost. Genes that were de-repressed lost or rearranged nucleosomes at their promoter, but not in the gene body. Therefore, a combination of DNA-encoded nucleosome stability and nucleosome composition dictates which nucleosomes will be lost under conditions of limiting histone protein. This, in turn, governs which genes will experience a loss of regulatory fidelity.
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| Overall design |
MNase-seq experiments consist of three wildtype (1 single-end and 2 paired-end) and four mutant (DCB200.1/H3 shutoff; 2 single-end, 2 paired-end) replicates. Each replicate contains two timepoints reflecting chromatin immediately after ("O hours") and 3 hours after transition to media containing dextrose. RNA-seq data includes three replicates from wildtype or H3 depleted cells after 3 hours in media containing dextrose.
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| Contributor(s) |
Gossett AJ, Lieb JD |
| Citation(s) |
22737086 |
| |
| Submission date |
May 14, 2011 |
| Last update date |
May 15, 2019 |
| Contact name |
Jason D Lieb |
| E-mail(s) |
[email protected]
|
| Phone |
919-843-3228
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| URL |
http://www.bio.unc.edu/faculty/lieb/labpages/default.shtml
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| Organization name |
University of North Carolina at Chapel Hill
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| Department |
Biology
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| Lab |
Jason Lieb
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| Street address |
203 Fordham Hall
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| City |
Chapel Hill |
| State/province |
NC |
| ZIP/Postal code |
27599 |
| Country |
USA |
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| Platforms (1) |
| GPL13272 |
Illumina Genome Analyzer IIx (Saccharomyces cerevisiae) |
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| Samples (8)
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| This SubSeries is part of SuperSeries: |
| GSE29294 |
Effects of Histone H3 depletion on nucleosome occupancy and positioning through the S. cerevisiae genome |
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| Relations |
| SRA |
SRP006797 |
| BioProject |
PRJNA142813 |
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