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SRX23902386: GSM8141295: WT H3K56ac eSPAN rep3; Schizosaccharomyces pombe; OTHER
1 ILLUMINA (NextSeq 500) run: 27.8M spots, 2.3G bases, 924.5Mb downloads

External Id: GSM8141295_r1
Submitted by: Irving Cancer Research Center, Pediatric and Genetics and Development, Columbia University
Study: Coordination of histone chaperones for parental histone segregation and epigenetic inheritance
show Abstracthide Abstract
Chromatin-based epigenetic memory relies on the accurate distribution of parental histone tetramers to newly replicated DNA strands, serving as templates for chromatin structure duplication. Mcm2, a subunit of the replicative helicase, and the Dpb3/4, subunits of polymerase epsilon, govern parental histone deposition to the lagging and leading strands, respectively. However, their contribution to epigenetic inheritance remains controversial. Here we show in fission yeast that a Mcm2 histone chaperone mutation severely disrupts heterochromatin inheritance, while Dpb3/4 mutations cause only moderate defects. Surprisingly, simultaneous Mcm2 and Dpb3/4 mutations stabilizes heterochromatin inheritance. eSPAN analyses confirm the conservation of Mcm2 and Dpb3/4 functions in parental histone segregation, with their collective absence reducing segregation bias. Furthermore, the FACT histone chaperone also regulates parental histone transfer independently of strands and collaborates with Mcm2 and Dpb3/4 to maintain parental histone density, ensuring faithful heterochromatin inheritance. These results underscore the importance of precise parental histone segregation to the lagging strand for epigenetic inheritance and unveil unique properties of parental histone chaperones during DNA replication. Overall design: Input, BrdU, H3K56ac ChIP-Seq, H3K56ac eSPAN, H3K4me3 ChIP-Seq, H3K4me3 eSPAN, are performed in S.pombe followed by Next Generation Sequencing
Sample: WT H3K56ac eSPAN rep3
SAMN40380402 • SRS20725738 • All experiments • All runs
Library:
Name: GSM8141295
Instrument: NextSeq 500
Strategy: OTHER
Source: GENOMIC
Selection: other
Layout: PAIRED
Construction protocol: Cells were crosslinked at 60 minutes after temperature shift with the addition of 1% formaldehyde and incubated for 20 minutes with shaking at 25°C. Immunoprecipitation was performed as described in ChIP. Crosslinking was reversed by Chelex-100. Briefly, total and ChIP DNA were incubated at 100°C for 5 minutes and then immediately cooled on ice for 5 minutes. DNA was diluted ten times with BrdU IP buffer (PBS, 0.0625% Triton X-100(v/v)), and incubated with BrdU antibody (BD Bioscience 555627) for two hours at 4°C. Next, Sepharose Protein G beads (Cytiva 17-0618-01) were added and incubated for an additional one hour at 4°C. The beads were then washed three times with BrdU IP buffer and once with TE. Finally, DNA was eluted with TES and purified with the QIAGEN MinElute PCR Purification kit (Qiagen 28004). Accel­NGS 1S Plus DNA library kit (Swift Bioscience, 10096).
Runs: 1 run, 27.8M spots, 2.3G bases, 924.5Mb
Run# of Spots# of BasesSizePublished
SRR2829330827,778,9562.3G924.5Mb2024-04-17

ID:
32207410

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