Abstract
In most bacteria, chromosome segregation is driven by the ParABS system where the CTPase protein ParB loads at the parS site to trigger the formation of a large partition complex. Here, we present in vitro studies of the partition complex for Bacillus subtilis ParB, using single-molecule fluorescence microscopy and AFM imaging to show that transient ParB–ParB bridges are essential for forming DNA condensates. Molecular Dynamics simulations confirm that condensation occurs abruptly at a critical concentration of ParB and show that multimerization is a prerequisite for forming the partition complex. Magnetic tweezer force spectroscopy on mutant ParB proteins demonstrates that CTP hydrolysis at the N-terminal domain is essential for DNA condensation. Finally, we show that transcribing RNA polymerases can steadily traverse the ParB–DNA partition complex. These findings uncover how ParB forms a stable yet dynamic partition complex for chromosome segregation that induces DNA condensation and segregation while enabling replication and transcription.
Original language | English |
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Article number | gkad868 |
Pages (from-to) | 11856-11875 |
Journal | Nucleic acids research |
Volume | 51 |
Issue number | 21 |
DOIs | |
Publication status | Published - 2023 |
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Dive into the research topics of 'Dynamic ParB–DNA interactions initiate and maintain a partition condensate for bacterial chromosome segregation'. Together they form a unique fingerprint.Datasets
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Data underlying chapter 5 of PhD thesis: Bacterial chromosome organization by ParB proteins
Tišma, M. (Creator), TU Delft - 4TU.ResearchData, 23 Nov 2023
DOI: 10.4121/DC4A4E4C-3FFD-4DBC-9310-89CA7607A596
Dataset/Software: Dataset