Cell wall damage increases macromolecular crowding effects in the Escherichia coli cytoplasm

iScience, vol. 26, pp. 106367

Abstract

The intracellular milieu is crowded with biomacromolecules. Macromolecular crowding changes the interactions, diffusion, and conformations of biomacromolecules. Changes in intracellular crowding have been mostly ascribed to differences in biomacromolecule concentration. However, spatial organization of these molecules should play a significant role in crowding effects. Here, we find that cell wall damage causes increased crowding effects in the Escherichia coli cytoplasm. Using a genetically encoded macromolecular crowding sensor, we see that crowding effects in spheroplasts and penicillin-treated cells well surpass crowding effects obtained using hyperosmotic stress. The crowding increase is not because of osmotic pressure, cell shape, or volume changes and therefore not crowder concentration. Instead, a genetically encoded nucleic acid stain and a DNA stain show cytoplasmic mixing and nucleoid expansion, which could cause these increased crowding effects. Our data demonstrate that cell wall damage alters the biochemical organization in the cytoplasm and induces significant conformational changes in a probe protein.

Authors 3

1. Theodoros Pittas Aachen DWI - Leibniz Institute for Interactive Materials Institute of Technical and Macromolecular Chemistry

   RWTH Aachen University · DWI – Leibniz Institute for Interactive Materials

   Affiliation as printed

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, NRW, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 1, 52074 Aachen, NRW, Germany

2. Weiyan Zuo Aachen DWI - Leibniz Institute for Interactive Materials Institute of Technical and Macromolecular Chemistry

   RWTH Aachen University · DWI – Leibniz Institute for Interactive Materials

   Affiliation as printed

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, NRW, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 1, 52074 Aachen, NRW, Germany

3. Arnold J. Boersma corresponding Aachen DWI - Leibniz Institute for Interactive Materials

   DWI – Leibniz Institute for Interactive Materials · Utrecht University

   Affiliation as printed

   Cellular Protein Chemistry, Bijvoet Centre for Biomolecular Research, Faculty of Science, Utrecht University, Utrecht, the Netherlands

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstrasse 50, 52074 Aachen, NRW, Germany