A flat embedding method for transmission electron microscopy reveals an unknown mechanism of tetracycline
Journal article, 2021

Transmission electron microscopy of cell sample sections is a popular technique in microbiology. Currently, ultrathin sectioning is done on resin-embedded cell pellets, which consumes milli- to deciliters of culture and results in sections of randomly orientated cells. This is problematic for rod-shaped bacteria and often precludes large-scale quantification of morphological phenotypes due to the lack of sufficient numbers of longitudinally cut cells. Here we report a flat embedding method that enables observation of thousands of longitudinally cut cells per single section and only requires microliter culture volumes. We successfully applied this technique to Bacillus subtilis, Escherichia coli, Mycobacterium bovis, and Acholeplasma laidlawii. To assess the potential of the technique to quantify morphological phenotypes, we monitored antibiotic-induced changes in B. subtilis cells. Surprisingly, we found that the ribosome inhibitor tetracycline causes membrane deformations. Further investigations showed that tetracycline disturbs membrane organization and localization of the peripheral membrane proteins MinD, MinC, and MreB. These observations are not the result of ribosome inhibition but constitute a secondary antibacterial activity of tetracycline that so far has defied discovery.

Peptidoglycans

Cell shape

Plasma protein Z

Author

Michaela Wenzel

University of Amsterdam

Chalmers, Biology and Biological Engineering, Chemical Biology

Swammerdam Institute for Life Sciences

Marien P. Dekker

Vrije Universiteit Amsterdam

Biwen Wang

Swammerdam Institute for Life Sciences

Maroeska J. Burggraaf

University of Amsterdam

Wilbert Bitter

University of Amsterdam

Vrije Universiteit Amsterdam

Jan R.T. van Weering

Vrije Universiteit Amsterdam

Leendert W. Hamoen

Swammerdam Institute for Life Sciences

Communications Biology

23993642 (eISSN)

Vol. 4 1 306-

Subject Categories

Cell Biology

Cell and Molecular Biology

Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

DOI

10.1038/s42003-021-01809-8

PubMed

33686188

More information

Latest update

4/22/2021