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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Dec 31, 2022 |
| Title |
ATR kinase supports normal proliferation in the early S phase by preventing replication resource exhaustion [CRISPR-Cas9 gRNA screen] |
| Organism |
Mus musculus |
| Experiment type |
Other
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| Summary |
The ATR kinase, which coordinates cellular responses to DNA replication stress, is also essential for the proliferation of normal unstressed cells. Although its role in the replication stress response is well defined, the mechanisms by which ATR supports normal cell proliferation remain elusive. Here, we show that Atr is dispensable for the viability of G0-arrested naïve B cells. However, upon cytokine-induced proliferation, Atr-deficient B cells initiate DNA replication efficiently in early S phase, but by mid-S phase they display dNTP depletion, fork stalling, and replication failure. Nonetheless, productive DNA replication can be restored in Atr-deficient cells by pathways that suppress origin firing, such as downregulation of CDC7 and CDK1 kinase activities. Together, these findings indicate that ATR supports the proliferation of normal unstressed cells by tempering the pace of origin firing during the early S phase to avoid exhaustion of dNTPs and other replication factors.
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| Overall design |
We set up a genome-wide CRISPR-Cas9 screen targeting 18,424 different genes with 4-5 gRNAs each in v-abl transformed wild-type mouse B cells treated with the IC90 dose of 1 of 2 ATR inhibitors (VE821 or AZD6738) or a Chk1 inhibitor (LY2603618), in triplicates, for 6 days and compared gRNA representation to DMSO-treated controls.
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| Contributor(s) |
Menolfi D, Lee BJ, Wang Y, Zhang H, Zha S |
| Citation(s) |
37336885 |
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| Submission date |
Oct 02, 2022 |
| Last update date |
Sep 08, 2023 |
| Contact name |
Shan Zha |
| E-mail(s) |
sz2296@cumc.columbia.edu
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| Organization name |
Columbia University
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| Street address |
1130 Saint Nicholas Ave.
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| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10032 |
| Country |
USA |
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| Platforms (1) |
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| Samples (18)
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| GSM6613558 |
v-abl transformed B cells, DMSO control, run1, rep1 |
| GSM6613559 |
v-abl transformed B cells, DMSO control, run1, rep2 |
| GSM6613560 |
v-abl transformed B cells, DMSO control, run1, rep3 |
| GSM6613561 |
v-abl transformed B cells, ATRi VE821, run1, rep1 |
| GSM6613562 |
v-abl transformed B cells, ATRi VE821, run1, rep2 |
| GSM6613563 |
v-abl transformed B cells, ATRi VE821, run1, rep3 |
| GSM6613564 |
v-abl transformed B cells, DMSO control, run2, rep1 |
| GSM6613565 |
v-abl transformed B cells, DMSO control, run2, rep2 |
| GSM6613566 |
v-abl transformed B cells, DMSO control, run2, rep3 |
| GSM6613567 |
v-abl transformed B cells, ATRi VE821, run2, rep1 |
| GSM6613568 |
v-abl transformed B cells, ATRi VE821, run2, rep2 |
| GSM6613569 |
v-abl transformed B cells, ATRi VE821, run2, rep3 |
| GSM6613570 |
v-abl transformed B cells, ATRi AZD6738, run2, rep1 |
| GSM6613571 |
v-abl transformed B cells, ATRi AZD6738, run2, rep2 |
| GSM6613572 |
v-abl transformed B cells, ATRi AZD6738, run2, rep3 |
| GSM6613573 |
v-abl transformed B cells, Chk1i LY2603618, run2, rep1 |
| GSM6613574 |
v-abl transformed B cells, Chk1i LY2603618, run2, rep2 |
| GSM6613575 |
v-abl transformed B cells, Chk1i LY2603618, run2, rep3 |
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| This SubSeries is part of SuperSeries: |
| GSE212196 |
ATR kinase supports normal proliferation in the early S phase by preventing replication resource exhaustion. |
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| Relations |
| BioProject |
PRJNA886447 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE214643_gRNA_20201112.raw_count.xlsx |
6.0 Mb |
(ftp)(http) |
XLSX |
| GSE214643_gRNA_20220118.raw_count.xlsx |
8.5 Mb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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