Autophagy is Regulated by Mitochondrial Calcium Transporters NCLX and MCU

De Miranda Ramos, V.; Serna, J. D.; Vilas-Boas, E. A.; Cabral-Costa, J. V.; da Cunha, F. M.; Kowaltowski, A. J.

doi:10.1101/2023.03.17.533187

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bioRxiv · 10.1101/2023.03.17.533187

Autophagy is Regulated by Mitochondrial Calcium Transporters NCLX and MCU

De Miranda Ramos, V.·Serna, J. D.·Vilas-Boas, E. A.·Cabral-Costa, J. V.·da Cunha, F. M.·Kowaltowski, A. J.

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Abstract

Mitochondria shape intracellular Ca2+ signaling through the concerted activity of Ca2+ uptake via mitochondrial calcium uniporter, and efflux from by Na+/Ca2+ exchangers (NCLX). Here, we describe a novel relationship between NCLX, intracellular Ca2+, and autophagic activity. Conditions that stimulate autophagy in vivo and in vitro, such as caloric restriction and nutrient deprivation, upregulate NCLX expression in hepatic tissue and cells. Conversely, knockdown of NCLX impairs basal and starvation-induced autophagy. Similarly, acute inhibition of NCLX activity by CGP 37157 affects bulk and endoplasmic reticulum autophagy (ER-phagy) without significant impacts on mitophagy. Mechanistically, CGP 37157 inhibited the formation of FIP200 puncta and downstream autophagosome biogenesis. Inhibition of NCLX caused decreased cytosolic Ca2+ levels, and intracellular Ca2+ chelation similarly suppressed autophagy. Furthermore, chelation did not exhibit an additive effect on NCLX inhibition of autophagy, demonstrating that mitochondrial Ca2+ efflux regulates autophagy through the modulation of Ca2+ signaling. Collectively, our results show that the mitochondrial Ca2+ extrusion pathway through NCLX is an important regulatory node linking nutrient restriction and autophagy regulation.

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De Miranda Ramos, V., Serna, J. D., Vilas-Boas, E. A., Cabral-Costa, J. V., da Cunha, F. M., Kowaltowski, A. J.. 2023-03-18. Autophagy is Regulated by Mitochondrial Calcium Transporters NCLX and MCU. https://doi.org/10.1101/2023.03.17.533187

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