NCBI Logo
GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
          Go
Series GSE73668 Query DataSets for GSE73668
Status Public on Oct 08, 2015
Title Methylation of histone H4 lysine 20 by PR-Set7 ensures the integrity of late replicating sequence domains in Drosophila
Organism Drosophila melanogaster
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary The methylation state of lysine 20 on histone H4 (H4K20) has been linked to cell cycle progression, Origin Recognition Complex (ORC) binding, and replication origin regulation. Monomethylation of H4K20 (H4K20me1) is mediated by the cell cycle-regulated histone methyltransferase PR-Set7, which is essential for genome integrity and cell cycle progression. PR-Set7 depletion in mammalian cells results in defective S-phase progression and the accumulation of DNA damage, which could be partially attributed to a defect in pre-Replication Complex (pre-RC) formation and origin activity. However, these studies were limited to a handful of mammalian origins, and it remains unclear how PR-Set7 and H4K20 methylation impact the replication program on a genomic scale. Using Drosophila Kc167 cells, we employed genetic, cytological, and genomic approaches to better understand the role of PR-Set7 and H4K20 methylation in regulating DNA replication and governing genome stability. We find that depletion of Drosophila PR-Set7 and loss of H4K20me1 result in the accumulation of DNA damage and an ATR-dependent cell cycle arrest. The cell cycle arrest occurs during the second S-phase following loss of PR-Set7 activity, suggesting that accumulation of nascent H4K20 is recalcitrant to the DNA replication program. Deregulation of H4K20 methylation had no impact on origin activation throughout the genome; instead, we found that the DNA damage marker, phosphorylated H2A.v (γ-H2A.v), accumulated specifically in late replicating domains in the absence of PR-Set7. This suggests that the molecular basis for the cell cycle arrest and accumulation of DNA damage resulting from loss of PR-Set7 is stochastic fork collapse within late replicating domains.
 
Overall design Examination of BrdU enrichment in the presence of hydroxyurea to map activation of early replication origin under control, PR-Set7, ATR, and ATR/PR-Set7 RNAi treatment conditions in duplicates using Illumina HiSeq and Miseq respectively; map early origin in 6 other conditions using Illumina HiSeq; examination of H4K20me1 in early or late S-phase with two different experimental methods, using Hiseq; examination of a histone variant with control or PR-Set7 RNAi treatment, in duplicates, using Hiseq.
 
Contributor(s) Li Y, Belsky JA, MacAlpine DM
Citation(s) 27131378
Submission date Oct 01, 2015
Last update date May 15, 2019
Contact name Jason Alan Belsky
E-mail(s) jason.belsky@duke.edu
Organization name Duke University
Department Pharmacology and Cancer Biology
Lab David MacAlpine Lab
Street address LSRC Room C333
City Durham
State/province NC
ZIP/Postal code 27710
Country USA
 
Platforms (2)
GPL13304 Illumina HiSeq 2000 (Drosophila melanogaster)
GPL16479 Illumina MiSeq (Drosophila melanogaster)
Samples (22)
GSM1900395 control_RNAi_rep1
GSM1900396 PR-Set7_RNAi_rep1
GSM1900397 ATR_RNAi_rep1
Relations
BioProject PRJNA297597
SRA SRP064393

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE73668_RAW.tar 18.9 Mb (http)(custom) TAR (of CSV)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

| NLM | NIH | GEO Help | Disclaimer | Accessibility |
NCBI Home NCBI Search NCBI SiteMap