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ING2 tumor suppressive protein translocates into mitochondria and is involved in cellular metabolism homeostasis

Abstract

ING2 (Inhibitor of Growth 2) is a tumor suppressor gene that has been implicated in critical biological functions (cell-cycle regulation, replicative senescence, DNA repair and DNA replication), most of which are recognized hallmarks of tumorigenesis occurring in the cell nucleus. As its close homolog ING1 has been recently observed in the mitochondrial compartment, we hypothesized that ING2 could also translocate into the mitochondria and be involved in new biological functions. In the present study, we demonstrate that ING2 is imported in the inner mitochondrial fraction in a redox-sensitive manner in human cells and that this mechanism is modulated by 14-3-3η protein expression. Remarkably, ING2 is necessary to maintain mitochondrial ultrastructure integrity without interfering with mitochondrial networks or polarization. We observed an interaction between ING2 and mtDNA under basal conditions. This interaction appears to be mediated by TFAM, a critical regulator of mtDNA integrity. The loss of mitochondrial ING2 does not impair mtDNA repair, replication or transcription but leads to a decrease in mitochondrial ROS production, suggesting a detrimental impact on OXPHOS activity. We finally show using multiple models that ING2 is involved in mitochondrial respiration and that its loss confers a protection against mitochondrial respiratory chain inhibition in vitro. Consequently, we propose a new tumor suppressor role for ING2 protein in the mitochondria as a metabolic shift gatekeeper during tumorigenesis.

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Fig. 1: ING2 is located in the inner mitochondrial compartment in human cell lines.
Fig. 2: Mitochondrial import of ING2 is redox-sensitive and is impaired upon inhibition of mitochondrial disulfide relay system.
Fig. 3: ING2 down-regulation impairs mitochondrial ultrastructure without interferring with the mitochondrial network.
Fig. 4: ING2 protein interacts with transcription factor A mitochondrial (TFAM) and is involved in the control of mitochondrial ROS production.
Fig. 5: ING2 loss impairs mitochondrial oxidative phosphorylation and decreases the cytotoxic effect of mitochondrial respiratory chain inhibitors.

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Acknowledgements

We thank S.Manié for his participation and the technical support. We thank the photonic facility of the Microscopy Rennes Imaging Center (MRic-Photonics) of Biosit, Université de Rennes 1. We thank Pr JA MacDonald and his team members as well as Pr K Riabowol (University of Calgary, Canada) for providing lentiviral vectors and technical support for the generation of the CrispR clones. AM was a recipient of a doctoral fellowship from La Ligue Contre le Cancer and Region Bretagne. C. Ricordel was a recipient of FHU Camin (CHU Rennes) doctoral followship and Nuovo-Soldati Fundation research grant. RP is supported by INSERM (Institut National de la Santé et de la Recherche Médicale). The work was supported by La Ligue Contre le Cancer (Grand Ouest), Association pour la Recherche sur le Cancer (ARC), Fondation pour la Recherche Médicale (FMR, DEQ20180339169), AIS Rennes Métropole, Biosit and Action Incitative Université de Rennes 1.

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CR, LC, AB, and RP conceived and designed the experiments. CR, AM, AB, AJ, LC, FJ, MT, and ABu performed the experiments. CR, LC, MT, and RP analyzed the data. RP, HL, BD, TG approved the manuscript and contributed to the financial support. CR and RP wrote the paper.

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Correspondence to Rémy Pedeux.

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Ricordel, C., Chaillot, L., Blondel, A. et al. ING2 tumor suppressive protein translocates into mitochondria and is involved in cellular metabolism homeostasis. Oncogene 40, 4111–4123 (2021). https://doi.org/10.1038/s41388-021-01832-3

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