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Visualization of Submitochondrial Protein Distributions

Apoptotic effectors in the intermembrane mitochondrial space – insights by STED nanoscopy

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By allowing visualization of submitochondrial protein distributions, fluorescence nanoscopy offers significant advantages for understanding cell death control. How cells manage, and control mitochondrial intermembrane proteins involved in apoptosis, such as Cytochrome c and DIABLO/SMAC is not completely defined. In this study, we used two-color STED nanoscopy to gain insight into the submitochondrial distribution of Cytochrome c and DIABLO/SMAC, two apoptotic effectors. Proteotoxic stress was used to induce cell death in leiomyosarcoma cells. STED microscopy allowed to reveal a cluster organization of DIABLO/SMAC distribution compared to the uniform distribution of ATP synthase. These clusters sometimes co-localize with clusters containing Cytochrome c. In conclusion, within the intermembrane space of mitochondria different storages of pro-death effectors exist.

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Iuliano L., Drioli S., Pignochino Y., Cafiero C.M., Minisini M., D'Este F., Picco R., Dalla E., Giordano G., Grignani G., Di Giorgio E., Benedetti F., Felluga F. and Brancolini C.:

Enhancing Proteotoxic Stress in Leiomyosarcoma Cells Triggers Mitochondrial Dysfunctions, Cell Death, and Antitumor Activity in vivo

DOI: 10.1158/1535-7163.MCT-20-0521 Published June 2021

https://mct.aacrjournals.org/content/early/2021/04/28/1535-7163.MCT-20-0521

Figure 1: Two-color STED imaging of submitochondrial distribution of Cytochrome c and DIABLO/SMAC. Image left: DIABLO/SMAC is organized in clusters compared to the uniform distribution of ATP synthase. Image right: Cytochrome c also shows clustered distribution and it sometimes co-localizes with clusters of DIABLO/SMAC. Images are processed with LASX LIGHTNING.

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