Dynamic ParB–DNA interactions initiate and maintain a partition condensate for bacterial chromosome segregation

M. Tišma, R. Janissen, Hammam Antar, A. Martin Gonzalez, R. Barth, T.G.T. Beekman, J. van der Torre, Davide Michieletto, Stephan Gruber, C. Dekker*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

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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 languageEnglish
Article numbergkad868
Pages (from-to)11856-11875
JournalNucleic acids research
Volume51
Issue number21
DOIs
Publication statusPublished - 2023

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