Home LiteratureArticle Details
PMID: 16157596 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

Suppression of hypoxia-inducible factor 1alpha (HIF-1alpha) transcriptional activity by the HIF prolyl hydroxylase EGLN1.

The Journal of biological chemistry ·Vol. 280 ·No. 45 ·2005-11-11 ·Pages 38102-7

To KK, Huang LE

Abstract

The cellular response to hypoxia is, at least in part, mediated by the transcriptional regulation of hypoxia-responsive genes involved in balancing the intracellular ATP production and consumption. Recent evidence suggests that the transcription factor, HIF-1alpha, functions as a master regulator of oxygen homeostasis by controlling a broad range of cellular events in hypoxia. In normoxia, HIF-1alpha is targeted for destruction via prolyl hydroxylation, an oxygen-dependent modification that signals for recognition by the ubiquitin ligase complex containing the von Hippel-Lindau tumor suppressor. Three HIF prolyl hydroxylases (EGLN1, EGLN2, and EGLN3) have been identified in mammals, among which EGLN1 and EGLN3 are hypoxia-inducible at their mRNA levels in an HIF-1alpha-dependent manner. In this study, we demonstrated that apart from promoting HIF-1alpha proteolysis in normoxia, EGLN1 specifically represses HIF-1alpha transcriptional activity in hypoxia. Ectopic expression of EGLN1 inhibited HIF-1alpha transcriptional activity without altering its protein levels in a von Hippel-Lindau-deficient cell line, indicating a discrete activity of EGLN1 in transcriptional repression. Conversely, silencing of EGLN1 expression augmented HIF-1alpha transcriptional activity and its target gene expression in hypoxia. Thus, we proposed that the accumulated EGLN1 in hypoxia acts as a negative-feedback mechanism to modulate HIF-1alpha target gene expression. Our finding also provided new insight into the pharmacological manipulation of the HIF prolyl hydroxylase for ischemic diseases.

MeSH Terms
Animals Cell Line Gene Silencing Genes, Reporter/genetics Humans Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors,genetics,metabolism Hypoxia-Inducible Factor-Proline Dioxygenases Immediate-Early Proteins/genetics,metabolism Oxygen/metabolism,pharmacology Procollagen-Proline Dioxygenase/genetics,metabolism Protein Binding/drug effects Transcription, Genetic
Chemicals
Hypoxia-Inducible Factor 1, alpha Subunit Immediate-Early Proteins EGLN1 protein, human Procollagen-Proline Dioxygenase Hypoxia-Inducible Factor-Proline Dioxygenases Oxygen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
To Kenneth K W
Laboratory of Human Carcinogenesis, NCI, National Institutes of Health, Bethesda, MD 20892, USA.
Huang L Eric
References (51)
51 references, click to expand
  1. Analysis of gene function in somatic mammalian cells using small interfering RNAs.
    Methods. 2002 Feb;26(2):199-213 PMID: 12054897
  2. Asparagine hydroxylation of the HIF transactivation domain a hypoxic switch.
    Science. 2002 Feb 1;295(5556):858-61 PMID: 11823643
  3. Hypoxia-inducible factor (HIF) asparagine hydroxylase is identical to factor inhibiting HIF (FIH) and is related to the cupin structural family.
    J Biol Chem. 2002 Jul 19;277(29):26351-5 PMID: 12042299
  4. Cellular adaptation to hypoxia: O2-sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression.
    FASEB J. 2002 Aug;16(10):1151-62 PMID: 12153983
  5. Hsp90 regulates a von Hippel Lindau-independent hypoxia-inducible factor-1 alpha-degradative pathway.
    J Biol Chem. 2002 Aug 16;277(33):29936-44 PMID: 12052835
  6. Leu-574 of HIF-1alpha is essential for the von Hippel-Lindau (VHL)-mediated degradation pathway.
    J Biol Chem. 2002 Nov 1;277(44):41750-5 PMID: 12205091
  7. Structure of factor-inhibiting hypoxia-inducible factor (HIF) reveals mechanism of oxidative modification of HIF-1 alpha.
    J Biol Chem. 2003 Jan 17;278(3):1802-6 PMID: 12446723
  8. Intracellular localisation of human HIF-1 alpha hydroxylases: implications for oxygen sensing.
    J Cell Sci. 2003 Apr 1;116(Pt 7):1319-26 PMID: 12615973
  9. Hypoxia-inducible factor and its biomedical relevance.
    J Biol Chem. 2003 May 30;278(22):19575-8 PMID: 12639949
  10. Regulation of angiogenesis by hypoxia: role of the HIF system.
    Nat Med. 2003 Jun;9(6):677-84 PMID: 12778166
  11. HIF prolyl-hydroxylase 2 is the key oxygen sensor setting low steady-state levels of HIF-1alpha in normoxia.
    EMBO J. 2003 Aug 15;22(16):4082-90 PMID: 12912907
  12. Targeting HIF-1 for cancer therapy.
    Nat Rev Cancer. 2003 Oct;3(10):721-32 PMID: 13130303
  13. HIF-1 as a target for drug development.
    Nat Rev Drug Discov. 2003 Oct;2(10):803-11 PMID: 14526383
  14. The von Hippel Lindau/hypoxia-inducible factor (HIF) pathway regulates the transcription of the HIF-proline hydroxylase genes in response to low oxygen.
    J Biol Chem. 2003 Dec 5;278(49):48690-5 PMID: 14506252
  15. HIF-1 and hypoxic response: the plot thickens.
    Curr Opin Genet Dev. 2004 Feb;14(1):81-5 PMID: 15108809
  16. HIF-1alpha induces cell cycle arrest by functionally counteracting Myc.
    EMBO J. 2004 May 5;23(9):1949-56 PMID: 15071503
  17. Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation.
    FASEB J. 2004 Jun;18(9):1028-30 PMID: 15084514
  18. The HIF pathway as a therapeutic target.
    Drug Discov Today. 2004 Aug 15;9(16):704-11 PMID: 15341784
  19. Differential function of the prolyl hydroxylases PHD1, PHD2, and PHD3 in the regulation of hypoxia-inducible factor.
    J Biol Chem. 2004 Sep 10;279(37):38458-65 PMID: 15247232
  20. Characterization of hypoxia-inducible factor 1 and regulation of DNA binding activity by hypoxia.
    J Biol Chem. 1993 Oct 15;268(29):21513-8 PMID: 8408001
  21. Purification and characterization of hypoxia-inducible factor 1.
    J Biol Chem. 1995 Jan 20;270(3):1230-7 PMID: 7836384
  22. Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension.
    Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5510-4 PMID: 7539918
  23. Dimerization, DNA binding, and transactivation properties of hypoxia-inducible factor 1.
    J Biol Chem. 1996 Jul 26;271(30):17771-8 PMID: 8663540
  24. Oxygen sensing and molecular adaptation to hypoxia.
    Physiol Rev. 1996 Jul;76(3):839-85 PMID: 8757790
  25. An essential role for p300/CBP in the cellular response to hypoxia.
    Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12969-73 PMID: 8917528
  26. Activation of hypoxia-inducible transcription factor depends primarily upon redox-sensitive stabilization of its alpha subunit.
    J Biol Chem. 1996 Dec 13;271(50):32253-9 PMID: 8943284
  27. Activation of hypoxia-inducible factor-1; definition of regulatory domains within the alpha subunit.
    J Biol Chem. 1997 Apr 25;272(17):11205-14 PMID: 9111021
  28. Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes.
    J Biol Chem. 1997 Sep 5;272(36):22642-7 PMID: 9278421
  29. Regulation of hypoxia-inducible factor 1alpha is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway.
    Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):7987-92 PMID: 9653127
  30. Signal transduction in hypoxic cells: inducible nuclear translocation and recruitment of the CBP/p300 coactivator by the hypoxia-inducible factor-1alpha.
    EMBO J. 1998 Nov 16;17(22):6573-86 PMID: 9822602
  31. Functional role of p35srj, a novel p300/CBP binding protein, during transactivation by HIF-1.
    Genes Dev. 1999 Jan 1;13(1):64-75 PMID: 9887100
  32. The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis.
    Nature. 1999 May 20;399(6733):271-5 PMID: 10353251
  33. Dynamic balancing of the dual nature of HIF-1alpha for cell survival.
    Cell Cycle. 2004 Jul;3(7):853-4 PMID: 15190211
  34. OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha.
    Mol Cell. 2005 Feb 18;17(4):503-12 PMID: 15721254
  35. HIF-1alpha induces genetic instability by transcriptionally downregulating MutSalpha expression.
    Mol Cell. 2005 Mar 18;17(6):793-803 PMID: 15780936
  36. The candidate tumor suppressor ING4 represses activation of the hypoxia inducible factor (HIF).
    Proc Natl Acad Sci U S A. 2005 May 24;102(21):7481-6 PMID: 15897452
  37. Regulation of mammalian O2 homeostasis by hypoxia-inducible factor 1.
    Annu Rev Cell Dev Biol. 1999;15:551-78 PMID: 10611972
  38. Ubiquitination of hypoxia-inducible factor requires direct binding to the beta-domain of the von Hippel-Lindau protein.
    Nat Cell Biol. 2000 Jul;2(7):423-7 PMID: 10878807
  39. Hypoxia inducible factor-alpha binding and ubiquitylation by the von Hippel-Lindau tumor suppressor protein.
    J Biol Chem. 2000 Aug 18;275(33):25733-41 PMID: 10823831
  40. Mechanism of regulation of the hypoxia-inducible factor-1 alpha by the von Hippel-Lindau tumor suppressor protein.
    EMBO J. 2000 Aug 15;19(16):4298-309 PMID: 10944113
  41. Activation of HIF1alpha ubiquitination by a reconstituted von Hippel-Lindau (VHL) tumor suppressor complex.
    Proc Natl Acad Sci U S A. 2000 Sep 12;97(19):10430-5 PMID: 10973499
  42. HIF-1-dependent transcriptional activity is required for oxygen-mediated HIF-1alpha degradation.
    FEBS Lett. 2001 Feb 23;491(1-2):85-90 PMID: 11226425
  43. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing.
    Science. 2001 Apr 20;292(5516):464-8 PMID: 11292862
  44. Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation.
    Science. 2001 Apr 20;292(5516):468-72 PMID: 11292861
  45. Molecular mechanism of hypoxia-inducible factor 1alpha -p300 interaction. A leucine-rich interface regulated by a single cysteine.
    J Biol Chem. 2001 Feb 2;276(5):3550-4 PMID: 11063749
  46. HIF-1alpha binding to VHL is regulated by stimulus-sensitive proline hydroxylation.
    Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9630-5 PMID: 11504942
  47. Characterization and comparative analysis of the EGLN gene family.
    Gene. 2001 Sep 5;275(1):125-32 PMID: 11574160
  48. C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation.
    Cell. 2001 Oct 5;107(1):43-54 PMID: 11595184
  49. FIH-1: a novel protein that interacts with HIF-1alpha and VHL to mediate repression of HIF-1 transcriptional activity.
    Genes Dev. 2001 Oct 15;15(20):2675-86 PMID: 11641274
  50. A conserved family of prolyl-4-hydroxylases that modify HIF.
    Science. 2001 Nov 9;294(5545):1337-40 PMID: 11598268
  51. FIH-1 is an asparaginyl hydroxylase enzyme that regulates the transcriptional activity of hypoxia-inducible factor.
    Genes Dev. 2002 Jun 15;16(12):1466-71 PMID: 12080085
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M504342200
Published
2005-11-11
Epub
2005-00-12
Pages
38102-7
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC1307502
Subset
IM
Grants
NCI NIH HHS · Z01 BC010424-06 · United States
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1074/jbc.M504342200 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2