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| Status |
Public on Dec 06, 2023 |
| Title |
Primase excites the competition of templating transcription and lagging replication to boost DNA damage |
| Organism |
Arabidopsis thaliana |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
Other
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| Summary |
Head-on transcription-replication conflicts (HO-TRCs) cause R-loops and DNA damage. We realized that HO-TRCs are the natural competition for templating transcription and lagging replication in the same DNA strand (termed TemLag competition), however, the regulatory mechanisms behind are unclear. We previously identified a chloroplast-localized RNase H1 protein AtRNH1C that can remove R-loops and relax HO-TRCs for genome integrity. Through the mutagenesis screen, we identified that the chloroplast-localized primase ATH exacerbates TemLag competition in the atrnh1c mutant. A mutation in ATH weakens the binding affinity of DNA, thus slowing down DNA replication and relieving TemLag competition. Overexpression of ATH boosts TemLag competition and intensifies DNA damage. Interestingly, strand-specific DNA damage sequencing reveals that TemLag competition induces single-strand DNA damage of the competitive strand at the end of transcription. These results illustrated a potentially conserved mechanism among organisms, of which the primase antagonizes R-loop clearance machinery to exaggerate TemLag competition and genome instability.
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| Overall design |
Detail-seq performed using chloroplasts embedded in low-melting-point agarose. ssDRIP performed with S9.6 antibody in atrnh1c mutant. ATH ChIP performed with GFP antibody using ATH-GFP tagged transgenic plants, and Col-0 with no tag as control.
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| Contributor(s) |
Zhang W, Yang Z, Wang W, Sun Q |
| Citation(s) |
38168108 |
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| Submission date |
Oct 13, 2022 |
| Last update date |
Jun 17, 2024 |
| Contact name |
Weifeng Zhang |
| Organization name |
Tsinghua University
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| Street address |
Zhongguancun Street
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| City |
Beijing |
| ZIP/Postal code |
100084 |
| Country |
China |
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| Platforms (1) |
| GPL26208 |
Illumina NovaSeq 6000 (Arabidopsis thaliana) |
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| Samples (16)
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| Relations |
| BioProject |
PRJNA890279 |
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