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Series GSE134665 Query DataSets for GSE134665
Status Public on Jul 22, 2020
Title Structural and biochemical analyses of Caulobacter crescentus ParB reveal the role of its N-terminal domain in chromosome segregation
Organisms Escherichia coli; Caulobacter vibrioides NA1000
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary The tripartite ParA-ParB-parS complex ensures faithful chromosome segregation in the majority of bacterial species. ParB nucleates on the centromere-like parS site and spreads to neighboring DNA to form a network of protein-DNA complexes. This nucleoprotein network in turn interacts with ParA to partition the parS locus, hence the chromosome to each daughter cell. Here, we determine the co-crystal structure of the C-terminal domain truncated ParB-parS complex from Caulobacter crescentus, and show that its N-terminal domain is inherently flexible and adopts multiple different conformations. We propose that the flexibility of the N-terminal domain might facilitate the spreading of ParB on the chromosome. Next, using ChIP-seq we show that ParBs from different bacterial species exhibit variation in their intrinsic capability for spreading, and that the N-terminal domain rather than the C-terminal domain is the main determinant for the variation in spreading. Finally, we show that the C-terminal domain of Caulobacter ParB does not possess non-specific DNA-binding activity in vitro. Engineered ParB variants with enhanced non-specific DNA-binding activity condense DNA in vitro but do not spread further than a wild-type protein in vivo. Taken together, our results emphasize the central role of the N-terminal domain in ParB spreading and faithful chromosome segregation.
Overall design Chromatin-immunoprecipitation with deep sequencing experiments (ChIP-seq) were performed on exponential-growing Escherichia coli and Caulobacter crescentus.
Contributor(s) Le TB, Tran NT, Jalal AS, Stevenson SE, Lawson DM, Moreno-Herrero F, Pastrana CL
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Submission date Jul 22, 2019
Last update date Jul 25, 2020
Contact name Tung Ba Khanh Le
Phone 01603450776
Organization name John Innes Centre
Department Department of Molecular Microbiology
Street address Colney Lane
City Norwich
State/province Norfolk
ZIP/Postal code NR4 7UH
Country United Kingdom
Platforms (2)
GPL18133 Illumina HiSeq 2500 (Escherichia coli)
GPL26943 Illumina HiSeq 2500 (Caulobacter vibrioides NA1000)
Samples (27)
GSM3963011 LeLab_ChIP_seq_MT148_vanA_Pvan_1xFLAG_Caulobacter_crescentus_ParB_3K_rep1
GSM3963012 LeLab_ChIP_seq_MT148_vanA_Pvan_1xFLAG_Caulobacter_crescentus_ParB_3K_rep2
GSM3963013 LeLab_ChIP_seq_MT148_vanA_Pvan_1xFLAG_Caulobacter_crescentus_ParB_4K_rep1
BioProject PRJNA556011
SRA SRP215947

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Supplementary file Size Download File type/resource
GSE134665_RAW.tar 310.0 Mb (http)(custom) TAR (of TXT)
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Raw data are available in SRA
Processed data provided as supplementary file

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