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| Status |
Public on Sep 08, 2021 |
| Title |
A regulatory phosphosite on Mec1 controls RNAPII and RNAPIII occupancy during replication stress [ChIP-seq] |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
Upon replication stress, the Mec1ATR kinase triggers the downregulation of transcription, reducing the level of RNA polymerase on chromatin to facilitate replication fork progression. We identify a hydroxyurea-induced phosphorylation site at Mec1-S1991 that contributes to the eviction of RNAPII and RNAPIII during replication stress. The non-phosphorylatable mec1-S1991A mutant reduces replication fork progression genome-wide and compromises survival on hydroxyurea. This defect can be suppressed by destabilizing chromatin-bound RNAPII with a Rpb3-TAP fusion, suggesting that lethality arises from replication-transcription conflicts. Coincident with a failure to repress gene expression, highly transcribed genes like GAL1 persist at nuclear pores in mec1-S1991A cells. Consistently, we find pore proteins and several components controlling RNAPII and RNAPIII transcription are phosphorylated in a Mec1-dependent manner, suggesting that Mec1-S1991 phosphorylation limits conflicts between replication and either RNA polymerase complex. We further show that Mec1 contributes to reduced RNAPII occupancy on chromatin during an unperturbed S phase.
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| Overall design |
Pol2 genome-wide profiling during DNA replication stress in wt-cells and mec1 mutants
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| |
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| Contributor(s) |
Jonas F, Barkai N |
| Citation(s) |
34569643 |
| Submission date |
Jul 15, 2021 |
| Last update date |
Nov 18, 2021 |
| Contact name |
romain forey |
| E-mail(s) |
romain.forey@epfl.ch
|
| Organization name |
IGH
|
| Street address |
142 avenue de la Cardonille
|
| City |
Montpellier |
| ZIP/Postal code |
34000 |
| Country |
France |
| |
|
| Platforms (2) |
| GPL19756 |
Illumina NextSeq 500 (Saccharomyces cerevisiae) |
| GPL27812 |
Illumina NovaSeq 6000 (Saccharomyces cerevisiae) |
|
| Samples (8)
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| This SubSeries is part of SuperSeries: |
| GSE180167 |
A regulatory phosphosite on Mec1 controls RNAPII and RNAPIII occupancy during replication stress |
|
| Relations |
| BioProject |
PRJNA746947 |
| SRA |
SRP328489 |
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