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Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression

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Abstract

Alteration of gene expression is a crucial component of adaptive responses to hypoxia. These responses are mediated by hypoxia-inducible transcription factors (HIFs)1,2. Here we describe an inhibitory PAS (Per/Arnt/Sim) domain protein, IPAS, which is a basic helix-loop-helix (bHLH)/PAS protein structurally related to HIFs. IPAS contains no endogenous transactivation function but demonstrates dominant negative regulation of HIF-mediated control of gene expression. Ectopic expression of IPAS in hepatoma cells selectively impairs induction of genes involved in adaptation to a hypoxic environment, notably the vascular endothelial growth factor (VEGF) gene, and results in retarded tumour growth and tumour vascular density in vivo. In mice, IPAS was predominantly expressed in Purkinje cells of the cerebellum and in corneal epithelium of the eye. Expression of IPAS in the cornea correlates with low levels of expression of the VEGF gene under hypoxic conditions. Application of an IPAS antisense oligonucleotide to the mouse cornea induced angiogenesis under normal oxygen conditions, and demonstrated hypoxia-dependent induction of VEGF gene expression in hypoxic corneal cells. These results indicate a previously unknown mechanism for negative regulation of angiogenesis and maintenance of an avascular phenotype.

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Figure 1: IPAS is a bHLH/PAS factor.
Figure 2: In situ hybridization analysis of IPAS expression in mouse cornea, retina and cerebellum.
Figure 3: IPAS is a dominant negative regulator of HIFs.
Figure 4: Expression of IPAS in hepatoma cells retards tumour growth.
Figure 5: Evidence that IPAS functions as an inhibitor of angiogenesis in the cornea.

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Acknowledgements

We thank Y. Fujii-Kuriyama and I. Pongratz for reagents. This work was supported by grants from the Swedish Medical Research Council, Pharmacia Corporation, Swedish Cancer Society, the Human Frontiers Science Program, the Uehara Memorial Foundation, and the Japanese Ministry of Education, Culture, Sports, Science and Technology.

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Author notes

  1. Mikael Asman & Anders Berkenstam

    Present address: Karobia AB, S-141 57, Huddinge, Sweden

  2. Göran Bertilsson

    Present address: Neuronova AB, S-114 33, Stockholm, Sweden

  3. Kristian Svensson

    Present address: Leica Microsystems AB, S-191 27, Solléntuna, Sweden

Authors and Affiliations

  1. Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institutet, Stockholm, S-171 77, Sweden

    Yuichi Makino & Lorenz Poellinger

  2. Department of Microbiology and Tumor Biology, Karolinska Institutet, Stockholm, S-171 77, Sweden

    Renhai Cao & Yihai Cao

  3. Center for Genomics and Bioinformatics, Karolinska Institutet, Stockholm, S-171 77, Sweden

    Mikael Asman

  4. Division of Clinical Immunology, Advanced Clinical Research Center, Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, 108-8639, Tokyo, Japan

    Yuichi Makino & Hirotoshi Tanaka

  5. Pharmacia Corporation, Stockholm, S-112 87, Sweden

    Kristian Svensson, Göran Bertilsson & Anders Berkenstam

  6. Karobia AB, S-141 57, Huddinge, Sweden

    Kristian Svensson & Göran Bertilsson

  7. Neuronova AB, S-114 33, Stockholm, Sweden

    Kristian Svensson & Anders Berkenstam

  8. Leica Microsystems AB, S-191 27, Solléntuna, Sweden

    Göran Bertilsson & Anders Berkenstam

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  1. Yuichi Makino
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  2. Renhai Cao
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  7. Yihai Cao
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  8. Anders Berkenstam
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  9. Lorenz Poellinger
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Correspondence to Lorenz Poellinger.

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Makino, Y., Cao, R., Svensson, K. et al. Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression. Nature 414, 550–554 (2001). https://doi.org/10.1038/35107085

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  • DOI: https://doi.org/10.1038/35107085

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