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ATR controls cellular adaptation to hypoxia through positive regulation of hypoxia-inducible factor 1 (HIF-1) expression

Oncogene volume 32, pages 4387–4396 (2013)Cite this article

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Abstract

Tumor cells adaptation to severe oxygen deprivation (hypoxia) plays a major role in tumor progression. The transcription factor HIF-1 (hypoxia-inducible factor 1), whose α-subunit is stabilized under hypoxic conditions is a key component of this process. Recent studies showed that two members of the phosphoinositide 3-kinase-related kinases (PIKKs) family, ATM (ataxia telangiectasia mutated) and DNA-PK (DNA-dependent protein kinase), regulate the hypoxic-dependent accumulation of HIF-1. These proteins initiate cellular stress responses when DNA damage occurs. In addition, it has been demonstrated that extreme hypoxia induces a replicative stress resulting in regions of single-stranded DNA at stalled replication forks and the activation of ATR (ataxia telangiectasia and Rad3 related protein), another member of the PIKKs family. Here, we show that even less severe hypoxia (0.1% O2) also induces activation of ATR through replicative stress. Importantly, in using either transiently silenced ATR cells, cells expressing an inactive form of ATR or cells exposed to an ATR inhibitor (CGK733), we demonstrate that hypoxic ATR activation positively regulates the key transcription factor HIF-1 independently of the checkpoint kinase Chk1. We show that ATR kinase activity regulates HIF-1α at the translational level and we find that the elements necessary for the regulation of HIF-1α translation are located within the coding region of HIF-1α mRNA. Finally, by using three independent cellular models, we clearly show that the loss of ATR expression and/or kinase activity results in the decrease of HIF-1 DNA binding under hypoxia and consequently affects protein expression levels of two HIF-1 target genes, GLUT-1 and CAIX. Taken together, our data show a new function for ATR in cellular adaptation to hypoxia through regulation of HIF-1α translation. Our work offers new prospect for cancer therapy using ATR inhibitors with the potential to decrease cellular adaptation in hypoxic tumors.

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Acknowledgements

We thank Catherine Botanch, Emilie Mirey and Christine Bordier for their help, Dr Nathalie Mazure and Dr Delphine Larrieu for critical comments on the manuscript and Aymeric Poizot for English rereading. We acknowledge Dr P Calsou, Dr Y Martineau and Dr S Pyronnet for technical advices and reagents. This work was supported by the ‘Ligue Nationale Contre le Cancer’ (Equipe Labélisée, BS), the Cancéropole GSO (BS and CM), the Association contre le Cancer, ARC (2008-1102, CM) and the Radioprotection Committee of EDF (CM).

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  1. B Salles

    Present address: 3Current address: TOXALIM, INRA, 180 chemin de Tournefeuille, 31027 Toulouse, France.,

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  1. CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), Toulouse, France

    F Fallone, S Britton, L Nieto, B Salles & C Muller

  2. Université de Toulouse, UPS, IPBS, Toulouse, France

    F Fallone, S Britton, L Nieto, B Salles & C Muller

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Correspondence to C Muller.

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Fallone, F., Britton, S., Nieto, L. et al. ATR controls cellular adaptation to hypoxia through positive regulation of hypoxia-inducible factor 1 (HIF-1) expression. Oncogene 32, 4387–4396 (2013). https://doi.org/10.1038/onc.2012.462

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