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

Miloš Tišma, Richard Janissen, Hammam Antar, Alejandro Martin-Gonzalez, Roman Barth, Twan Beekman, Jaco Van Der Torre, Davide Michieletto, Stephan Gruber, Cees Dekker*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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
Pages (from-to)11856-11875
Number of pages20
JournalNucleic Acids Research
Volume51
Issue number21
Early online date18 Oct 2023
DOIs
Publication statusPublished - 27 Nov 2023

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