Plasma membrane perforation by GSDME during apoptosis‑driven secondary necrosis

Elke De Schutter, Jana Ramon, Benjamin Pfeuty, Caroline De Tender, Stephan Stremersch, Koen Raemdonck, Wim Declercq, Franck Riquet, Kevin Braeckmans, Peter Vandenabeele

Research output: Contribution to journalA1: Web of Science-articlepeer-review

Abstract

Secondary necrosis has long been perceived as an uncontrolled process resulting in total lysis of the apoptotic cell. Recently, it was shown that progression of apoptosis to secondary necrosis is regulated by Gasdermin E (GSDME), which requires activation by caspase-3. Although the contribution of GSDME in this context has been attributed to its pore-forming capacity, little is known about the kinetics and size characteristics of this. Here we report on the membrane permeabilizing features of GSDME by monitoring the influx and efflux of dextrans of different sizes into/from anti-Fas-treated L929sAhFas cells undergoing apoptosis-driven secondary necrosis. We found that GSDME accelerates cell lysis measured by SYTOX Blue staining but does not affect the exposure of phosphatidylserine on the plasma membrane. Furthermore, loss of GSDME expression clearly hampered the influx of fluorescently labeled dextrans while the efflux happened independently of the presence or absence of GSDME expression. Importantly, both in- and efflux of dextrans were dependent on their molecular weight. Altogether, our results demonstrate that GSDME regulates the passage of compounds together with other plasma membrane destabilizing subroutines.
Original languageEnglish
Article number19 (2022)
JournalCellular and molecular life sciences
Volume79
ISSN1420-682X
DOIs
Publication statusPublished - 31-Dec-2021

Keywords

  • Animals
  • Apoptosis
  • Cell Line
  • Cell Membrane Permeability
  • Cell Membrane/metabolism
  • Cell Nucleus/metabolism
  • Dextrans/metabolism
  • Kinetics
  • Mice
  • Molecular Weight
  • Nanoparticles/chemistry
  • Necrosis/metabolism
  • Receptors, Estrogen/metabolism

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