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| Status |
Public on Sep 07, 2019 |
| Title |
Distinct transcriptional roles for Histone H3-K56 acetylation during the cell cycle |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
Dynamic disruption and reassembly of promoter-proximal nucleosomes is a conserved hallmark of transcriptionally active chromatin. Histone H3-K56 acetylation (H3K56Ac) enhances these turnover events and promotes nucleosome assembly during S phase. Here we sequence nascent transcripts to investigate the impact of H3K56Ac on transcription throughout the yeast cell cycle. Strikingly, we find that H3K56Ac is a genome-wide activator of transcription. H3K56Ac has a major impact on transcription initiation, but it also appears to promote elongation and/or termination. In contrast, H3K56Ac represses promiscuous transcription that occurs immediately following replication fork passage, in this case by promoting efficient nucleosome assembly. We also detect a stepwise increase in transcription as cells transit S phase and enter G2, but this response to increased gene dosage does not require H3K56Ac. Thus, a single histone mark can exert both positive and negative impacts on transcription that are coupled to different cell cycle events.
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| Overall design |
We have mapped changes in nucleosome positions on newly replicated DNA in a timecourse after genome replication. We have used Micrococcal Nuclease footprinting of cross linked chromatin to determine nucleosome positions and EdU (ethylene deoxy uridine) to mark nascent DNA strands. EdU incorporated into nascent DNA strands was biotinylated with Click chemistry and nascent DNA strand fragments were subsequently isolated using Streptavidin coated magnetic beads.
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| Contributor(s) |
Topal S, Vasseur P, Radman-Livaja M, Peterson CL |
| Citation(s) |
31558720 |
| Submission date |
Feb 18, 2019 |
| Last update date |
Oct 08, 2019 |
| Contact name |
Marta Radman-Livaja |
| E-mail(s) |
mrl5374@gmail.com
|
| Phone |
+33434359667
|
| Organization name |
CNRS
|
| Department |
IGMM
|
| Street address |
1919 route de Mende
|
| City |
Montpellier |
| ZIP/Postal code |
34293 |
| Country |
France |
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| Platforms (1) |
| GPL13821 |
Illumina HiSeq 2000 (Saccharomyces cerevisiae) |
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| Samples (13)
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| GSM3611236 |
EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [0 min after Thymidine chase] |
| GSM3611237 |
EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [2 min after Thymidine chase] |
| GSM3611238 |
EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [4 min after Thymidine chase] |
| GSM3611239 |
EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [6 min after Thymidine chase] |
| GSM3611240 |
EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [8 min after Thymidine chase] |
| GSM3611241 |
EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [15 min after Thymidine chase] |
| GSM3611242 |
Replicate of EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [2 min after Thymidine chase] |
| GSM3611243 |
Replicate of EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [4 min after Thymidine chase] |
| GSM3611244 |
Replicate of EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [6 min after Thymidine chase] |
| GSM3611245 |
Replicate of EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [8 min after Thymidine chase] |
| GSM3611246 |
Replicate of EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [15 min after Thymidine chase] |
| GSM3611247 |
Replicate of EdU pulse (5min) Thymidine chase (5min) input DNA time course rtt109D mutant [25 min after Thymidine chase] |
| GSM3611248 |
MNAse-seq of total chromatin in rtt109D mutant |
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| Relations |
| BioProject |
PRJNA522996 |
| SRA |
SRP186165 |
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