This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow E-mail this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Pan, Y.
Right arrow Articles by Simon, M. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Pan, Y.
Right arrow Articles by Simon, M. C.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, February 2007, p. 912-925, Vol. 27, No. 3
0270-7306/07/$08.00+0     doi:10.1128/MCB.01223-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Multiple Factors Affecting Cellular Redox Status and Energy Metabolism Modulate Hypoxia-Inducible Factor Prolyl Hydroxylase Activity In Vivo and In Vitro{triangledown}

Yi Pan,1,2,{dagger} Kyle D. Mansfield,2,{dagger},{ddagger} Cara C. Bertozzi,2 Viktoriya Rudenko,2 Denise A. Chan,3 Amato J. Giaccia,3 and M. Celeste Simon1,2*

Howard Hughes Medical Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104,1 Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104,2 Center for Clinical Science Research, Department of Radiation Oncology, Stanford University, Stanford, California 943053

Received 6 July 2006/ Returned for modification 4 August 2006/ Accepted 2 November 2006

Prolyl hydroxylation of hypoxible-inducible factor alpha (HIF-{alpha}) proteins is essential for their recognition by pVHL containing ubiquitin ligase complexes and subsequent degradation in oxygen (O2)-replete cells. Therefore, HIF prolyl hydroxylase (PHD) enzymatic activity is critical for the regulation of cellular responses to O2 deprivation (hypoxia). Using a fusion protein containing the human HIF-1{alpha} O2-dependent degradation domain (ODD), we monitored PHD activity both in vivo and in cell-free systems. This novel assay allows the simultaneous detection of both hydroxylated and nonhydroxylated PHD substrates in cells and during in vitro reactions. Importantly, the ODD fusion protein is regulated with kinetics identical to endogenous HIF-1{alpha} during cellular hypoxia and reoxygenation. Using in vitro assays, we demonstrated that the levels of iron (Fe), ascorbate, and various tricarboxylic acid (TCA) cycle intermediates affect PHD activity. The intracellular levels of these factors also modulate PHD function and HIF-1{alpha} accumulation in vivo. Furthermore, cells treated with mitochondrial inhibitors, such as rotenone and myxothiazol, provided direct evidence that PHDs remain active in hypoxic cells lacking functional mitochondria. Our results suggest that multiple mitochondrial products, including TCA cycle intermediates and reactive oxygen species, can coordinate PHD activity, HIF stabilization, and cellular responses to O2 depletion.

* Corresponding author. Mailing address: Howard Hughes Medical Institute, Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, 421 Curie Boulevard, Philadelphia, PA 19104. Phone: (215) 746-5532. Fax: (215) 746-5511. E-mail: celeste2{at}mail.med.upenn.edu.

{triangledown} Published ahead of print on 13 November 2006.

{dagger} Y.P. and K.D.M. contributed equally to this study.

{ddagger} Present address: Molecular Genetics and Microbiology, Duke University, Durham, NC 27710.

Molecular and Cellular Biology, February 2007, p. 912-925, Vol. 27, No. 3
0270-7306/07/$08.00+0     doi:10.1128/MCB.01223-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wiczer, B. M., Bernlohr, D. A. (2009). A novel role for fatty acid transport protein 1 in the regulation of tricarboxylic acid cycle and mitochondrial function in 3T3-L1 adipocytes. J. Lipid Res. 50: 2502-2513 [Abstract] [Full Text]  
  • Shao, Z., Zhang, Y., Powell-Coffman, J. A. (2009). Two Distinct Roles for EGL-9 in the Regulation of HIF-1-Mediated Gene Expression in Caenorhabditis elegans. Genetics 183: 821-829 [Abstract] [Full Text]  
  • Lee, W. H., Kim, Y. W., Choi, J. H., Brooks, S. C. III, Lee, M.-O., Kim, S. G. (2009). Oltipraz and dithiolethione congeners inhibit hypoxia-inducible factor-1{alpha} activity through p70 ribosomal S6 kinase-1 inhibition and H2O2-scavenging effect. Molecular Cancer Therapeutics 8: 2791-2802 [Abstract] [Full Text]  
  • Liu, Y., Veena, C. K., Morgan, J. B., Mohammed, K. A., Jekabsons, M. B., Nagle, D. G., Zhou, Y.-D. (2009). Methylalpinumisoflavone Inhibits Hypoxia-inducible Factor-1 (HIF-1) Activation by Simultaneously Targeting Multiple Pathways. J. Biol. Chem. 284: 5859-5868 [Abstract] [Full Text]  
  • Milovanova, T. N., Bhopale, V. M., Sorokina, E. M., Moore, J. S., Hunt, T. K., Hauer-Jensen, M., Velazquez, O. C., Thom, S. R. (2009). Hyperbaric oxygen stimulates vasculogenic stem cell growth and differentiation in vivo. J. Appl. Physiol. 106: 711-728 [Abstract] [Full Text]  
  • Aprelikova, O., Pandolfi, S., Tackett, S., Ferreira, M., Salnikow, K., Ward, Y., Risinger, J. I., Barrett, J. C., Niederhuber, J. (2009). Melanoma Antigen-11 Inhibits the Hypoxia-Inducible Factor Prolyl Hydroxylase 2 and Activates Hypoxic Response. Cancer Res. 69: 616-624 [Abstract] [Full Text]  
  • Mylonis, I., Chachami, G., Paraskeva, E., Simos, G. (2008). Atypical CRM1-dependent Nuclear Export Signal Mediates Regulation of Hypoxia-inducible Factor-1{alpha} by MAPK. J. Biol. Chem. 283: 27620-27627 [Abstract] [Full Text]  
  • Qutub, A. A., Popel, A. S. (2008). Reactive Oxygen Species Regulate Hypoxia-Inducible Factor 1{alpha} Differentially in Cancer and Ischemia. Mol. Cell. Biol. 28: 5106-5119 [Abstract] [Full Text]  
  • McFate, T., Mohyeldin, A., Lu, H., Thakar, J., Henriques, J., Halim, N. D., Wu, H., Schell, M. J., Tsang, T. M., Teahan, O., Zhou, S., Califano, J. A., Jeoung, N. H., Harris, R. A., Verma, A. (2008). Pyruvate Dehydrogenase Complex Activity Controls Metabolic and Malignant Phenotype in Cancer Cells. J. Biol. Chem. 283: 22700-22708 [Abstract] [Full Text]  
  • Park, Y.-K., Ahn, D.-R., Oh, M., Lee, T., Yang, E. G., Son, M., Park, H. (2008). Nitric Oxide Donor, ({+/-})-S-Nitroso-N-acetylpenicillamine, Stabilizes Transactive Hypoxia-Inducible Factor-1{alpha} by Inhibiting von Hippel-Lindau Recruitment and Asparagine Hydroxylation. Mol. Pharmacol. 74: 236-245 [Abstract] [Full Text]  
  • Zhang, H., Bosch-Marce, M., Shimoda, L. A., Tan, Y. S., Baek, J. H., Wesley, J. B., Gonzalez, F. J., Semenza, G. L. (2008). Mitochondrial Autophagy Is an HIF-1-dependent Adaptive Metabolic Response to Hypoxia. J. Biol. Chem. 283: 10892-10903 [Abstract] [Full Text]  
  • Jung, S.-N., Yang, W. K., Kim, J., Kim, H. S., Kim, E. J., Yun, H., Park, H., Kim, S. S., Choe, W., Kang, I., Ha, J. (2008). Reactive oxygen species stabilize hypoxia-inducible factor-1 alpha protein and stimulate transcriptional activity via AMP-activated protein kinase in DU145 human prostate cancer cells. Carcinogenesis 29: 713-721 [Abstract] [Full Text]  
  • Page, E. L., Chan, D. A., Giaccia, A. J., Levine, M., Richard, D. E. (2008). Hypoxia-inducible Factor-1{alpha} Stabilization in Nonhypoxic Conditions: Role of Oxidation and Intracellular Ascorbate Depletion. Mol. Biol. Cell 19: 86-94 [Abstract] [Full Text]  
  • Ferguson, J. E. III, Wu, Y., Smith, K., Charles, P., Powers, K., Wang, H., Patterson, C. (2007). ASB4 Is a Hydroxylation Substrate of FIH and Promotes Vascular Differentiation via an Oxygen-Dependent Mechanism. Mol. Cell. Biol. 27: 6407-6419 [Abstract] [Full Text]  
  • Clanton, T. L. (2007). Hypoxia-induced reactive oxygen species formation in skeletal muscle. J. Appl. Physiol. 102: 2379-2388 [Abstract] [Full Text]  


Copyright © 2010 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»