Reconstituting bacterial cell division assemblies in crowded, phase-separated media

Methods Enzymol. 2021:646:19-49. doi: 10.1016/bs.mie.2020.06.012. Epub 2020 Jul 9.

Abstract

Here we have summarized several strategies to reconstruct complexes containing the FtsZ protein, a central element of the cell division machinery in most bacteria, and to test their functional organization in minimal membrane systems and cell-like containers, as vesicles and droplets produced by microfluidics. These synthetic systems have been devised to mimic elements of the intracellular complexity, as excluded volume effects due to natural crowding, and macromolecular condensation resulting from biologically regulated liquid-liquid phase separation, in media of known and controllable composition. This integrative approach has allowed to demonstrate that macromolecular phase separation and crowding may also help to dynamically organize FtsZ in the intracellular space thus modulating its functional reactivity in cell division.

Keywords: Bimolecular condensation; Biomimetic membranes; Bottom-up synthetic biology; Cellular biochemistry; Liquid-liquid phase separation; Macromolecular crowding; Microfluidics; Minimal FtsZ-based divisomes; Molecular interactions; Self-organization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria*
  • Bacterial Proteins / genetics
  • Cell Division
  • Macromolecular Substances
  • Microfluidics*

Substances

  • Bacterial Proteins
  • Macromolecular Substances