|
| Status |
Public on Jun 23, 2022 |
| Title |
The ParB clamp docks onto Smc for DNA loading via a joint-ParB interface [ChIP-seq] |
| Organism |
Bacillus subtilis |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
Chromosomes readily unlink from one another and segregate to daughter cells during cell division highlighting a remarkable ability of cells to organize long DNA molecules. SMC complexes mediate chromosome folding by DNA loop extrusion. In most bacteria, SMC complexes start loop extrusion at the ParB/parS partition complex formed near the replication origin. Whether they are recruited by recognizing a specific DNA structure in the partition complex or a protein component is unknown. By replacing genes in Bacillus subtilis with orthologous sequences from Streptococcus pneumoniae, we show that the three subunits of the bacterial Smc complex together with the ParB protein form a functional module that can organize and segregate chromosomes when transplanted into another organism. Using chimeric proteins and chemical cross-linking, we find that ParB binds to the Smc subunit directly. We map a binding interface to the Smc joint and the ParB CTP-binding domain. Structure prediction indicates how the ParB clamp presents DNA to the Smc complex to initiate DNA loop extrusion.
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| |
|
| Overall design |
Chromatin-immunoprecipitation (ChIP) sequencing of ScpB in Bacillus subtilis ParB variants
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| |
|
| Contributor(s) |
Liu HW, Bock FP, Gruber S |
| Citation(s) |
36044845 |
| |
| Submission date |
Jun 20, 2022 |
| Last update date |
Sep 22, 2022 |
| Contact name |
Hon-Wing Liu |
| E-mail(s) |
hon-wing.liu@unil.ch
|
| Phone |
0766963933
|
| Organization name |
Université de Lausanne
|
| Street address |
Le Biophore, CH-1015 Lausanne
|
| City |
Ecublens VD |
| ZIP/Postal code |
1025 |
| Country |
Switzerland |
| |
|
| Platforms (1) |
| GPL30886 |
Illumina NovaSeq 6000 (Bacillus subtilis) |
|
| Samples (9)
|
| GSM6254207 |
Input DNA of strain BSG5146 (WT ParB) |
| GSM6254208 |
Input DNA of strain BSG5149 (parB(L69S,K70E,E101K)) |
| GSM6254209 |
Input DNA of strain BSG1050 (∆parB) |
| GSM6254210 |
ScpB ChIP DNA of strain BSG5146 (WT ParB) repeat 1 |
| GSM6254211 |
ScpB ChIP DNA of strain BSG5149 (parB(L69S,K70E,E101K)) repeat 1 |
| GSM6254212 |
ScpB ChIP DNA of strain BSG1050 (∆parB) repeat 1 |
| GSM6254213 |
ScpB ChIP DNA of strain BSG5146 (WT ParB) repeat 2 |
| GSM6254214 |
ScpB ChIP DNA of strain BSG5149 (parB(L69S,K70E,E101K)) repeat 2 |
| GSM6254215 |
ScpB ChIP DNA of strain BSG1050 (∆parB) repeat 2 |
|
| Relations |
| BioProject |
PRJNA850938 |
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