HIF1α stabilization in hypoxia is not oxidant-initiated

  1. Amit Kumar
  2. Manisha Vaish
  3. Saravanan S Karuppagounder
  4. Irina Gazaryan
  5. John W Cave
  6. Anatoly A Starkov
  7. Elizabeth T Anderson
  8. Sheng Zhang
  9. John T Pinto
  10. Austin M Rountree
  11. Wang Wang
  12. Ian R Sweet
  13. Rajiv R Ratan  Is a corresponding author
  1. Burke Neurological Institute, United States
  2. Regeneron Pharmaceuticals Inc, United States
  3. New York Medical College, United States
  4. InVitro Cell Research, United States
  5. Weill Medical College of Cornell University, United States
  6. Cornell university, United States
  7. University of Washington, United States

Abstract

Hypoxic adaptation mediated by HIF transcription factors requires mitochondria, which have been implicated in regulating HIF1α stability in hypoxia by distinct models that involve consuming oxygen or alternatively converting oxygen into the second messenger peroxide. Here, we use a ratiometric, peroxide reporter, HyPer to evaluate the role of peroxide in regulating HIF1α stability. We show that antioxidant enzymes are neither homeostatically induced nor are peroxide levels increased in hypoxia. Additionally, forced expression of diverse antioxidant enzymes, all of which diminish peroxide, had disparate effects on HIF1α protein stability. Moreover, decrease in lipid peroxides by glutathione peroxidase-4 or superoxide by mitochondrial SOD, failed to influence HIF1α protein stability. These data show that mitochondrial, cytosolic or lipid ROS were not necessary for HIF1α stability, and favor a model where mitochondria contribute to hypoxic adaptation as oxygen consumers.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file.

Article and author information

Author details

  1. Amit Kumar

    Neuroscience, Burke Neurological Institute, White Plains, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5017-9887
  2. Manisha Vaish

    Muscle and Metabolism, Regeneron Pharmaceuticals Inc, Tarrytown, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Saravanan S Karuppagounder

    Neuroscience, Burke Neurological Institute, White Plains, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Irina Gazaryan

    Department of Anatomy and Cell Biology, New York Medical College, Valhalla, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. John W Cave

    Neuroscience, InVitro Cell Research, Englewood, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Anatoly A Starkov

    Brain and Mind Research Institute and Department of Neurology, Weill Medical College of Cornell University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Elizabeth T Anderson

    Cornell university, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Sheng Zhang

    Cornell university, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8206-1007
  9. John T Pinto

    Department of Anatomy and Cell Biology, New York Medical College, Valhalla, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Austin M Rountree

    University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Wang Wang

    University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Ian R Sweet

    University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7565-1663
  13. Rajiv R Ratan

    Neuroscience, Burke Neurological Institute, White Plains, United States
    For correspondence
    rrr2001@med.cornell.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9081-2701

Funding

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2021, Kumar et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,281
    views
  • 405
    downloads
  • 18
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Amit Kumar
  2. Manisha Vaish
  3. Saravanan S Karuppagounder
  4. Irina Gazaryan
  5. John W Cave
  6. Anatoly A Starkov
  7. Elizabeth T Anderson
  8. Sheng Zhang
  9. John T Pinto
  10. Austin M Rountree
  11. Wang Wang
  12. Ian R Sweet
  13. Rajiv R Ratan
(2021)
HIF1α stabilization in hypoxia is not oxidant-initiated
eLife 10:e72873.
https://doi.org/10.7554/eLife.72873

Share this article

https://doi.org/10.7554/eLife.72873

Further reading

    1. Cell Biology
    Jittoku Ihara, Yibin Huang ... Koichi Yamamoto
    Research Article

    Chronic kidney disease (CKD) and atherosclerotic heart disease, frequently associated with dyslipidemia and hypertension, represent significant health concerns. We investigated the interplay among these conditions, focusing on the role of oxidized low-density lipoprotein (oxLDL) and angiotensin II (Ang II) in renal injury via G protein αq subunit (Gq) signaling. We hypothesized that oxLDL enhances Ang II-induced Gq signaling via the AT1 (Ang II type 1 receptor)-LOX1 (lectin-like oxLDL receptor) complex. Based on CHO and renal cell model experiments, oxLDL alone did not activate Gq signaling. However, when combined with Ang II, it significantly potentiated Gq-mediated inositol phosphate 1 production and calcium influx in cells expressing both LOX-1 and AT1 but not in AT1-expressing cells. This suggests a critical synergistic interaction between oxLDL and Ang II in the AT1-LOX1 complex. Conformational studies using AT1 biosensors have indicated a unique receptor conformational change due to the oxLDL-Ang II combination. In vivo, wild-type mice fed a high-fat diet with Ang II infusion presented exacerbated renal dysfunction, whereas LOX-1 knockout mice did not, underscoring the pathophysiological relevance of the AT1-LOX1 interaction in renal damage. These findings highlight a novel mechanism of renal dysfunction in CKD driven by dyslipidemia and hypertension and suggest the therapeutic potential of AT1-LOX1 receptor complex in patients with these comorbidities.

    1. Cell Biology
    Qi Zeng, Chen Yao ... Shuai Chen
    Research Article

    Mounting evidence has demonstrated the genetic association of ORMDL sphingolipid biosynthesis regulator 3 (ORMDL3) gene polymorphisms with bronchial asthma and a diverse set of inflammatory disorders. However, its role in type I interferon (type I IFN) signaling remains poorly defined. Herein, we report that ORMDL3 is a negative modulator of the type I IFN signaling by interacting with mitochondrial antiviral signaling protein (MAVS) and subsequently promoting the proteasome-mediated degradation of retinoic acid-inducible gene I (RIG-I). Immunoprecipitation coupled with mass spectrometry (IP-MS) assays uncovered that ORMDL3 binds to ubiquitin-specific protease 10 (USP10), which forms a complex with and stabilizes RIG-I through decreasing its K48-linked ubiquitination. ORMDL3 thus disrupts the interaction between USP10 and RIG-I, thereby promoting RIG-I degradation. Additionally, subcutaneous syngeneic tumor models in C57BL/6 mice revealed that inhibition of ORMDL3 enhances anti-tumor efficacy by augmenting the proportion of cytotoxic CD8 positive T cells and IFN production in the tumor microenvironment (TME). Collectively, our findings reveal the pivotal roles of ORMDL3 in maintaining antiviral innate immune responses and anti-tumor immunity.