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Series GSE135102 Query DataSets for GSE135102
Status Public on Dec 01, 2019
Title The nucleosome core particle remembers its position through DNA replication and transcription
Organism Saccharomyces cerevisiae
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary We tracked the fate of labelled nucleosomes through DNA replication, and established that nucleosomes present at a locus remembered their position during DNA replication. The replication-associated histone chaperones DPB3 and MCM2 were essential for nucleosome position memory, and in the absence of both DPB3 and MCM2 histone chaperone activity nucleosomes did not remember their position. Using the same approach, we tested the model that transcription results in retrograde transposition of nucleosomes along a transcription unit. We found no evidence of retrograde transposition. Our results suggest that nucleosomes have the capacity to transmit epigenetic memory across mitotic generations with exquisite spatial fidelity.
 
Overall design Nucleosomes were tagged with the 15aa AviTag, then the AviTag was biotinylated using various versions of TetR-BirA (including a split TetR-BirA obligate heterodimer). Site-specific biotinyaltion was stopped by adding doxycycline, and the labeled nucleosomes were detected before and after replication using streptavidin affinity purification and sequencing. In the same samples, we directly detected the localization of the TetR-BirA enzyme by using a V5-tagged allele of TetR-BirA, and precipitating the TetR-BirA-bound chromatin using anti-V5 agarose resin.
 
Contributor(s) Schlissel GS, Rine J
Citation(s) 31511420
Submission date Jul 30, 2019
Last update date Jan 15, 2020
Contact name Gavin Schwebs Schlissel
E-mail(s) gschliss@gmail.com
Phone 9256428068
Organization name UC Berkeley
Department Molecular Cell Biology
Lab Rine
Street address 440 Barker Hall
City Berkeley
State/province CA
ZIP/Postal code 94720
Country USA
 
Platforms (2)
GPL21656 Illumina HiSeq 4000 (Saccharomyces cerevisiae)
GPL22715 Illumina MiniSeq (Saccharomyces cerevisiae)
Samples (81)
GSM3985930 GSTS_001: JRy12392_strepSeq_cycling
GSM3985931 GSTS_002: JRy12392_v5-ChIP-seq cycling
GSM3985932 GSTS_003: JRy12392_strepSeq_arrested_dox
Relations
BioProject PRJNA557450
SRA SRP216839

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Supplementary file Size Download File type/resource
GSE135102_RAW.tar 8.6 Gb (http)(custom) TAR (of BEDGRAPH)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file
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