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| Status |
Public on Jun 02, 2017 |
| Title |
Phosphorylated SIRT1 associates with replication origins to prevent excess replication initiation and preserve genomic stability |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
Other
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| Summary |
Chromatin structure affects DNA replication patterns, but the role of specific chromatin modifiers in regulating the replication process is yet unclear. We report that phosphorylation of the human SIRT1 deacetylase on Threonine 530 (T530-pSIRT1) modulates DNA synthesis. T530-pSIRT1 associates with replication origins and inhibits replication from a group of ÒdormantÓ potential replication origins, which initiate replication only when cells are subject to replication stress. Although both active and dormant origins bind T530-pSIRT1, active origins are distinguished from dormant origins by their unique association with an open chromatin mark, histone H3 methylated on lysine 4. SIRT1 phosphorylation also facilitates leading and lagging strand coordination. SIRT1 T530 phosphorylation is essential to prevent DNA breakage upon replication stress and cells harboring SIRT1 that cannot be phosphorylated exhibit a high prevalence of extrachromosomal elements, hallmarks of perturbed replication. These observations suggest that SIRT1 phosphorylation modulates the distribution of replication initiation events to insure genomic stability.
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| Overall design |
Nascent strands were purified with the lambda exonuclease methods from HCT116 cells, which untreated or was treated 200nM aphidicolin for 24hrs and released into freshmedium, then cultured another 24hrs and cell cycle fractionated K562. Chromatin from unsyncrhonized untreated cultures of HCT116 cells was subjected to ChIP-Seq with antibody directed against T530-pSIRT1, S139-pMCM2 and H4K16Ac. Chromatin from cell cycle fractionalted K562 cells are subject to ChIP-Seq with antibody directed against T530-pSIRT1 and S139-pMCM2
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| Contributor(s) |
Aladjem MI |
| Citation(s) |
28549174 |
| |
| Submission date |
Feb 02, 2017 |
| Last update date |
May 15, 2019 |
| Contact name |
Christophe E Redon |
| E-mail(s) |
redonc@mail.nih.gov
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| Phone |
240-760-7338
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| Organization name |
NIH
|
| Street address |
37 Convent Drive
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (1) |
| GPL10999 |
Illumina Genome Analyzer IIx (Homo sapiens) |
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| Samples (23)
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| Relations |
| BioProject |
PRJNA369608 |
| SRA |
SRP098727 |
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