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| Status |
Public on May 01, 2021 |
| Title |
A CTP-dependent gating mechanism enables ParB spreading on DNA |
| Organisms |
Escherichia coli; Caulobacter vibrioides |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
Proper chromosome segregation is essential in all living organisms. The ParA-ParB-parS system is widely employed for chromosome segregation in bacteria. Previously, we showed that Caulobacter crescentus ParB requires cytidine triphosphate to escape the nucleation site parS to spread by sliding to the neighboring DNA. Here, we provide the structural basis for this transition from nucleation to spreading by solving co-crystal structures of a C-terminal domain truncated C. crescentus ParB with parS and with a CTP analog. Nucleating ParB is an open clamp, in which parS is captured at the DNA-binding domain (the DNA-gate). Upon binding CTP, the N-terminal domain (NTD) self-dimerizes to close the NTD-gate of the clamp. The DNA-gate also closes, thus driving parS into a compartment between the DNA-gate and the C-terminal domain. CTP hydrolysis and/or the release of hydrolytic products may re-open the gates. Overall, we suggest a CTP-operated gating mechanism that regulates ParB nucleation and spreading.
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| Overall design |
Chromatin-immunoprecipitation with deep sequencing experiments (ChIP-seq) were performed on exponential-growing Caulobacter crescentus (see the supplementary Materials and Methods for the exact treatment that were applied to each strain).
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| Contributor(s) |
Jalal AS, Tran NT, Stevenson CE, Lawson DM, Le TB |
| Citation(s) |
34397383 |
| |
| Submission date |
Mar 15, 2021 |
| Last update date |
Sep 05, 2021 |
| Contact name |
Tung Ba Khanh Le |
| E-mail(s) |
tung.le@jic.ac.uk
|
| Phone |
01603450776
|
| Organization name |
John Innes Centre
|
| Department |
Department of Molecular Microbiology
|
| Lab |
www.tunglelab.org
|
| Street address |
Colney Lane
|
| City |
Norwich |
| State/province |
Norfolk |
| ZIP/Postal code |
NR4 7UH |
| Country |
United Kingdom |
| |
|
| Platforms (2) |
| GPL18006 |
Illumina HiSeq 2500 (Caulobacter vibrioides) |
| GPL18133 |
Illumina HiSeq 2500 (Escherichia coli) |
|
| Samples (8)
|
| GSM5173186 |
LELab ChIP seq TLS3050 anti FLAG replicate1 |
| GSM5173187 |
LELab ChIP seq TLS3050 anti FLAG replicate2 |
| GSM5173188 |
LELab ChIP seq TLS3059 anti FLAG replicate1 |
| GSM5173189 |
LELab ChIP seq TLS3059 anti FLAG replicate2 |
| GSM5462477 |
LELab_ChIP_seq_TLS3079_anti_GFP_replicate1 |
| GSM5462478 |
LELab_ChIP_seq_TLS3079_anti_GFP_replicate2 |
| GSM5462479 |
LELab_ChIP_seq_TLS3080_anti_GFP_replicate1 |
| GSM5462480 |
LELab_ChIP_seq_TLS3080_anti_GFP_replicate2 |
|
| Relations |
| BioProject |
PRJNA714673 |
| SRA |
SRP310754 |
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