Homeostatic mitochondrial lipid peroxidation governs mitochondrial fission site selection and daughter mitochondrial fate
Abstract
Mitochondrial fission is essential for cellular homeostasis, yet signals that license specific sites for division and link fission to subsequent fate remain largely elusive. Here, we identify homeostatic inner mitochondrial membrane lipid peroxidation (IMM LPO) as an upstream signal coordinating these two fundamental processes. We show, during homeostatic conditions, a focal IMM LPO increase is necessary for division, marking mitochondria-ER contacts (MERCS) for fission. IMM LPO and MERCS formation can precede one another, engaging in a positive feedback loop: focal IMM LPO promotes ER recruitment and subsequent mitochondrial Ca²⁺ influx, which in turn accelerates IMM LPO, creating a robust commitment for division. Critically, the magnitude of the IMM LPO signal at the prospective fission site dictates daughter mitochondrial fate. Modest LPO generates healthy daughter mitochondria, while severe LPO targets depolarized daughters for mitophagy. This IMM LPO-mediated fission is a general mechanism, exhibiting hyperactivation in prostate cancer. Our study defines a fundamental initiation event for mitochondrial fission, establishing homeostatic IMM LPO as a signal that integrates fission site selection with mitochondrial fate.
References
1. Youle, R.J. & van der Bliek, A.M. Mitochondrial fission, fusion, and stress. Science 337, 1062–1065 (2012).
2. Liu, L., Li, Y., Chen, G. & Chen, Q. Crosstalk between mitochondrial biogenesis and mitophagy to maintain mitochondrial homeostasis. J. Biomed. Sci. 30, 86 (2023).
3. Friedman, J.R. et al. ER tubules mark sites of mitochondrial division. Science 334, 358–362 (2011).
4. Steffen, J. & Koehler, C.M. ER–mitochondria contacts: Actin dynamics at the ER control mitochondrial fission via calcium release. J. Cell Biol. 217, 15–17 (2018).
5. Smirnova, E., Griparic, L., Shurland, D.-L. & van der Bliek, A.M. Dynamin-related protein Drp1 is required for mitochondrial division in mammalian cells. Mol. Biol. Cell 12, 2245–2256 (2001).
6. Fonseca, T.B., Sánchez-Guerrero, Á., Milosevic, I. & Raimundo, N. Mitochondrial fission requires DRP1 but not dynamins. Nature 570, E34–E42 (2019).
7. Lewis, S.C., Uchiyama, L.F. & Nunnari, J. ER-mitochondria contacts couple mtDNA synthesis with mitochondrial division in human cells. Science 353, aaf5549 (2016).
8. Tábara, L.-C., Segawa, M. & Prudent, J. Molecular mechanisms of mitochondrial dynamics. Nat. Rev. Mol. Cell Biol. 26, 123–146 (2025).
9. Twig, G. et al. Fission and selective fusion govern mitochondrial segregation and elimination by autophagy. EMBO J. 27, 433–446 (2008).
10. Tábara, L.C. et al. MTFP1 controls mitochondrial fusion to regulate inner membrane quality control and maintain mtDNA levels. Cell 187, 3619–3637 (2024).
11. Mishra, S.R. et al. PPA2 activates MTFP1-DNM1L fission signaling to govern mitochondrial proliferation and mitophagy. Autophagy 22, 121–144 (2026).
12. Kleele, T. et al. Distinct fission signatures predict mitochondrial degradation or biogenesis. Nature 593, 435–439 (2021).
13. Das, A.T., Tenenbaum, L. & Berkhout, B. Tet-on systems for doxycycline-inducible gene expression. Curr. Gene Ther. 16, 156–167 (2016).
14. Liu, Y., Wan, Y., Jiang, Y., Zhang, L. & Cheng, W. GPX4: The hub of lipid oxidation, ferroptosis, disease and treatment. Biochim. Biophys. Acta Rev. Cancer 1878, 188890 (2023).
15. Shioji, K., Oyama, Y., Okuma, K. & Nakagawa, H. Synthesis and properties of fluorescence probe for detection of peroxides in mitochondria. Bioorg. Med. Chem. Lett. 20, 3911–3915 (2010).
16. Tosatto, S.C. et al. The catalytic site of glutathione peroxidases. Antioxid. Redox Signal. 10, 1515–1526 (2008).
17. Taguchi, N., Ishihara, N., Jofuku, A., Oka, T. & Mihara, K. Mitotic phosphorylation of dynamin-related GTPase Drp1 participates in mitochondrial fission. J. Biol. Chem. 282, 11521–11529 (2007).
18. Chang, C.-R. & Blackstone, C. Cyclic AMP-dependent protein kinase phosphorylation of Drp1 regulates its GTPase activity and mitochondrial morphology. J. Biol. Chem. 282, 21583–21587 (2007).
19. Prime, T.A. et al. A ratiometric fluorescent probe for assessing mitochondrial phospholipid peroxidation within living cells. Free Radic. Biol. Med. 53, 544–553 (2012).
20. Wang, F. et al. PALP: A rapid imaging technique for stratifying ferroptosis sensitivity in normal and tumor tissues in situ. Cell Chem. Biol. 29, 157–170 (2022).
21. Abrisch, R.G., Gumbin, S.C., Wisniewski, B.T., Lackner, L.L. & Voeltz, G.K. Fission and fusion machineries converge at ER contact sites to regulate mitochondrial morphology. J. Cell Biol. 219, e201911122 (2020).
22. Mishima, E. et al. DHODH inhibitors sensitize to ferroptosis by FSP1 inhibition. Nature 619, E9–E18 (2023).
23. Cho, B. et al. Constriction of the mitochondrial inner compartment is a priming event for mitochondrial division. Nat. Commun. 8, 15754 (2017).
24. Erdahl, W.L., Chapman, C.J., Taylor, R.W. & Pfeiffer, D.R. Ca2+ transport properties of ionophores A23187, ionomycin, and 4-BrA23187 in a well defined model system. Biophys. J. 66, 1678–1693 (1994).
25. Bartok, A. et al. IP3 receptor isoforms differently regulate ER-mitochondrial contacts and local calcium transfer. Nat. Commun. 10, 3726 (2019).
26. Baughman, J.M. et al. Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature 476, 341–345 (2011).
27. Maruyama, T., Kanaji, T., Nakade, S., Kanno, T. & Mikoshiba, K. 2APB, 2-aminoethoxydiphenyl borate, a membrane-penetrable modulator of Ins(1,4,5)P3-induced Ca2+ release. J. Biochem. 122, 498–505 (1997).
28. Liu, Y. et al. MCU-induced mitochondrial calcium uptake promotes mitochondrial biogenesis and colorectal cancer growth. Signal Transduct. Target. Ther. 5, 59 (2020).
29. Blomme, A. et al. 2,4-dienoyl-CoA reductase regulates lipid homeostasis in treatment-resistant prostate cancer. Nat. Commun. 11, 2508 (2020).
30. Gao, D. et al. Acetate utilization promotes hormone therapy resistance in prostate cancer through neuroendocrine differentiation. Drug Resist. Updat. 77, 101158 (2024).
31. Xu, H. et al. Targeting RBP4-STRA6 retinol signaling disrupts adipose-prostate crosstalk: A novel strategy to suppress basal cell plasticity in androgen deprivation. Metabolism 169, 156288 (2025).
32. Dixon, S.J. et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell 149, 1060–1072 (2012).
33. Decker, S.T. & Funai, K. Mitochondrial membrane lipids in the regulation of bioenergetic flux. Cell Metab. 36, 1963–1978 (2024).
34. Yang, W.S. et al. Peroxidation of polyunsaturated fatty acids by lipoxygenases drives ferroptosis. Proc. Natl. Acad. Sci. USA 113, E4966–E4975 (2016).
35. Cartes-Saavedra, B., Ghosh, A. & Hajnóczky, G. The roles of mitochondria in global and local intracellular calcium signalling. Nat. Rev. Mol. Cell Biol. 26, 456–475 (2025).
36. Nolfi-Donegan, D., Braganza, A. & Shiva, S. Mitochondrial electron transport chain: Oxidative phosphorylation, oxidant production, and methods of measurement. Redox Biol. 37, 101674 (2020).
37. Angelova, P.R., Esteras, N. & Abramov, A.Y. Mitochondria and lipid peroxidation in the mechanism of neurodegeneration: Finding ways for prevention. Med. Res. Rev. 41, 770–784 (2021).
38. Golla, V.K., Boyd, K.J. & May, E.R. Curvature sensing lipid dynamics in a mitochondrial inner membrane model. Commun. Biol. 7, 29 (2024).
39. Benink, H.A. & Urh, M. HaloTag technology for specific and covalent labeling of fusion proteins. Methods Mol Biol 1266, 119–128 (2015).
40. Valente, A.J., Maddalena, L.A., Robb, E.L., Moradi, F. & Stuart, J.A. A simple ImageJ macro tool for analyzing mitochondrial network morphology in mammalian cell culture. Acta Histochem. 119, 315–326 (2017).
Metrics
DOI:
Submission ID:
Downloads
Additional Files
Supplemental File(s)
Posted
How to Cite
Download Citation
Declaration of Competing Interests
The authors declare no competing interests to disclose.
Copyright
The copyright holder for this preprint is the author/funder.
All rights reserved. This work is protected by copyright. No part of this work may be reproduced, distributed, or transmitted in any form or by any means without the prior written permission of the copyright holder.