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Cyclosporin A Prevents the Hypoxic Adaptation by Activating Hypoxia-inducible Factor-1α Pro-564 Hydroxylation

Abstract : The mechanism by which hypoxia induces gene transcription involves the inhibition of hypoxia-inducible factor (HIF)-1α prolyl hydroxylase activity, which prevents von Hippel-Lindau (vHL)-dependent targeting of HIF-1α to the ubiquitin-proteasome pathway. HIF-1α is stabilized, translocates to the nucleus, interacts with hypoxia-responsive elements, and promotes the activation of target genes. This report shows that cyclosporin A (CsA) interferes with the hypoxic signaling cascade in C6 glioma cells. CsA inhibits hypoxia-dependent gene transcription in a reporter gene assay and prevents the hypoxic accumulation of HIF-1α. Addition of the 530–603 C-terminal oxygen-dependent degradation (ODD) domain of HIF-1α to the green fluorescent protein (GFP) destabilized the protein in an oxygen-dependent manner. CsA prevented the hypoxic stabilization of an ODD·GFP fusion protein. An assay for 2-oxoglutarate-dependent dioxygenases was developed using a light mitochondrial kidney fraction as a source of enzyme. It uses the capacity of specific peptides to stimulate the degradation of [14C]2-oxoglutarate. CsA stimulated the enzymatic activity in the presence of a peptide that mimicked the 557–576 sequence of HIF-1α. The enzyme promoted [35S]vHL binding to glutathione S-transferase (GST)·ODD fusion protein. This association increased in the presence of CsA. CsA effects were not observed when the proline residue corresponding to Pro-564 in the HIF-1α sequence was replaced by a hydroxyproline or an alanine residue. Finally, CsA increased vHL-ODD interaction during hypoxia. We conclude that CsA destabilizes HIF-1α by promoting hydroxylation of Pro-564 in the ODD domain. Such a mechanism may prevent hypoxic adaptation during CsA-induced nephrotoxicity and contribute to the adverse effects of this drug.
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https://hal.archives-ouvertes.fr/hal-03034018
Contributor : Gisela D'ANGELO Connect in order to contact the contributor
Submitted on : Tuesday, November 9, 2021 - 4:12:32 PM
Last modification on : Thursday, August 4, 2022 - 4:52:50 PM
Long-term archiving on: : Thursday, February 10, 2022 - 7:21:59 PM

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Gisela d'Angelo, Eric Duplan, Paul Vigne, Christian Frelin. Cyclosporin A Prevents the Hypoxic Adaptation by Activating Hypoxia-inducible Factor-1α Pro-564 Hydroxylation. Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2003, 278 (17), pp.15406-15411. ⟨10.1074/jbc.m211293200⟩. ⟨hal-03034018⟩

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