Condensation and localization of the partitioning protein ParB on the bacterial chromosome

Proc Natl Acad Sci U S A. 2014 Jun 17;111(24):8809-14. doi: 10.1073/pnas.1402529111. Epub 2014 Jun 3.

Abstract

The ParABS system mediates chromosome segregation and plasmid partitioning in many bacteria. As part of the partitioning mechanism, ParB proteins form a nucleoprotein complex at parS sites. The biophysical basis underlying ParB-DNA complex formation and localization remains elusive. Specifically, it is unclear whether ParB spreads in 1D along DNA or assembles into a 3D protein-DNA complex. We show that a combination of 1D spreading bonds and a single 3D bridging bond between ParB proteins constitutes a minimal model for a condensed ParB-DNA complex. This model implies a scaling behavior for ParB-mediated silencing of parS-flanking genes, which we confirm to be satisfied by experimental data from P1 plasmids. Furthermore, this model is consistent with experiments on the effects of DNA roadblocks on ParB localization. Finally, we show experimentally that a single parS site is necessary and sufficient for ParB-DNA complex formation in vivo. Together with our model, this suggests that ParB binding to parS triggers a conformational switch in ParB that overcomes a nucleation barrier. Conceptually, the combination of spreading and bridging bonds in our model provides a surface tension ensuring the condensation of the ParB-DNA complex, with analogies to liquid-like compartments such as nucleoli in eukaryotes.

Keywords: par system; protein localization; protein-DNA condensate.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Algorithms
  • Bacterial Physiological Phenomena
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / physiology*
  • Binding Sites
  • Chromosomes, Bacterial / chemistry*
  • DNA, Bacterial / chemistry*
  • DNA-Binding Proteins / chemistry
  • Gene Silencing
  • Genomics
  • Green Fluorescent Proteins / chemistry
  • Kinetics
  • Monte Carlo Method
  • Plasmids / metabolism
  • Protein Binding
  • Protein Conformation
  • Software
  • Temperature

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • DNA-Binding Proteins
  • chromosome partition proteins, bacterial
  • Green Fluorescent Proteins