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| Status |
Public on Jan 18, 2018 |
| Title |
SLFN11 blocks stressed replication forks independently of ATR (NS-Seq) |
| Organism |
Homo sapiens |
| Experiment type |
Other
|
| Summary |
SLFN11 sensitizes cancer cells to a broad range of DNA-targeted therapies. Here we show that, in response to replication stress induced by camptothecin, SLFN11 tightly binds chromatin at stressed replication foci via RPA1 together with the replication helicase subunit MCM3. Unlike ATR, SLFN11 neither interferes with the loading of CDC45 and PCNA nor inhibits the initiation of DNA replication but selectively blocks fork progression while inducing chromatin opening across replication initiation sites. The ATPase domain of SLFN11 is required for chromatin opening, replication block and cell death but not for the tight binding of SLFN11 to chromatin. Replication stress by the CHK1 inhibitor Prexasertib also recruits SLFN11 to nascent replicating DNA together with CDC45 and PCNA. We conclude that SLFN11 is recruited to stressed replication forks carrying extended RPA filaments where it blocks replication by changing chromatin structure across replication sites.
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| Overall design |
Nascent strands were purified with the lambda exonuclease methods from CCRF-CEM parental and SLFN11-KO (knockout) cell lines, which were treated with DMSO, 100 nM camptothecine or 100 nM camptothecin plus 2 uM VE-821 (ATR inhibitor) for 4 hrs.
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| Contributor(s) |
Pommier Y, Murai J, Redon C |
| Citation(s) |
29395061 |
| |
| Submission date |
Jul 17, 2017 |
| Last update date |
Jul 25, 2021 |
| Contact name |
Junko Murai |
| E-mail(s) |
junko.murai@nih.gov
|
| Organization name |
NCI
|
| Street address |
37 Convent Dr.
|
| City |
Bethesda |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (1) |
| GPL10999 |
Illumina Genome Analyzer IIx (Homo sapiens) |
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| Samples (8)
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| GSM2705263 |
Nascent strands reads from CCRF-CEM parental cells Control |
| GSM2705264 |
Nascent strands reads from CCRF-CEM parental cells CPT |
| GSM2705265 |
Nascent strands reads from CCRF-CEM parental cells CPTATRi |
| GSM2705266 |
Nascent strands reads from CCRF-CEM SLFN11-KO cells control |
| GSM2705267 |
Nascent strands reads from CCRF-CEM SLFN11-KO cells CPT |
| GSM2705268 |
Nascent strands reads from CCRF-CEM SLFN11-KO cells CPTATRi |
| GSM2705269 |
Control Reads (random shear) from CCRF-CEM parental cells (NS-Seq) |
| GSM2705270 |
Control Reads (random shear) from CCRF-CEM SLFN11-KO (NS-Seq) |
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| This SubSeries is part of SuperSeries: |
| GSE101516 |
SLFN11 blocks stressed replication forks independently of ATR |
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| Relations |
| BioProject |
PRJNA394712 |
| SRA |
SRP112549 |
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