2023
bioRxiv

Endogenous BAX and BAK form mosaic rings of variable size and composition on apoptotic mitochondria

Authors:

Sarah Vanessa Schweighofer, Daniel C. Jans, Jan Keller-Findeisen, Anne Folmeg, Mark Bates, Stefan Jakobs

Keywords:

apoptosis, BAX, BAK, mitochondria, apoptotic pore

Abstract:

One hallmark of apoptosis is the oligomerization of BAX and BAK to form a pore in the mitochondrial outer membrane, which mediates the release of pro-apoptotic intermembrane space proteins into the cytosol. Cells overexpressing BAX or BAK fusion proteins are a powerful model system to study the dynamics and localization of these proteins in cells. However, it is unclear whether overexpressed BAX and BAK form the same ultrastructural assemblies following the same temporal hierarchy as endogenously expressed proteins. Combining live– and fixed-cell STED super-resolution microscopy, we show that overexpression of BAK results in novel BAK structures, which are virtually absent in non-overexpressing apoptotic cells. We further demonstrate that in wildtype cells, BAK is recruited to apoptotic pores before BAX. Both proteins together form unordered, mosaic rings on apoptotic mitochondria in immortalized cell culture models as well as in human primary cells. In BAX– or BAK-single-knockout cells, the remaining protein is able to form rings independently. The heterogeneous nature of these rings corroborates the toroidal apoptotic pore model.