Abstract
Hypoxia-inducible factor 1alpha (HIF-1alpha) is rapidly degraded by the ubiquitin-proteasome pathway under normoxic conditions. Ubiquitination of HIF-1alpha is mediated by interaction with von Hippel-Lindau tumor suppressor protein (pVHL). In our previous report, we found that hypoxia-induced active signal transducer and activator of transcription3 (STAT3) accelerated the accumulation of HIF-1alpha protein and prolonged its half-life in solid tumor cells. However, its specific mechanisms are not fully understood. Thus, we examined the role of STAT3 in the mechanism of pVHL-mediated HIF-1alpha stability. We found that STAT3 interacts with C-terminal domain of HIF-1alpha and stabilizes HIF-1alpha by inhibition of pVHL binding to HIF-1alpha. The binding between HIF-1alpha and pVHL, negative regulator of HIF-1alpha stability, was interfered dose-dependently by overexpressed constitutive active STAT3. Moreover, we found that the enhanced HIF-1alpha protein levels by active STAT3 are due to decrease of poly-ubiquitination of HIF-1alpha protein via inhibition of interaction between pVHL and HIF-1alpha. Taken together, our results suggest that STAT3 decreases the pVHL-mediated ubiquitination of HIF-1alpha through competition with pVHL for binding to HIF-1 alpha, and then stabilizes HIF-1alpha protein levels.
MeSH Terms
Animals
COS Cells
Cell Line, Tumor
Chlorocebus aethiops
Humans
Hypoxia-Inducible Factor 1, alpha Subunit/genetics,metabolism
Immunoblotting
Immunoprecipitation
Protein Binding
STAT3 Transcription Factor/genetics,metabolism
Signal Transduction/genetics,physiology
Transfection
Ubiquitination
Von Hippel-Lindau Tumor Suppressor Protein/genetics,metabolism
Chemicals
Hypoxia-Inducible Factor 1, alpha Subunit
STAT3 Transcription Factor
Von Hippel-Lindau Tumor Suppressor Protein
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Jung Joo Eun
Department of Pharmacology, Seoul National University College of Medicine, Seoul 110-799, Korea.
Kim Hong Sook
Lee Chang Seok
Shin Yong Jae
Kim Yong Nyun
Kang Gyeong Hoon
Kim Tae You
Juhnn Yong Sung
Kim Sung Joon
Park Jong Wan
Ye Sang Kyu
Chung Myung Hee
References (21)
21 references, click to expand
-
HIF hydroxylation and the mammalian oxygen-sensing pathway.
J Clin Invest. 2003 Mar;111(6):779-83
PMID: 12639980
-
Nur77 upregulates HIF-alpha by inhibiting pVHL-mediated degradation.
Exp Mol Med. 2008 Feb 29;40(1):71-83
PMID: 18305400
-
STAT signaling in head and neck cancer.
Oncogene. 2000 May 15;19(21):2489-95
PMID: 10851047
-
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
-
Constitutive activation of stat 3 oncogene product in human ovarian carcinoma cells.
Gynecol Oncol. 2000 Oct;79(1):67-73
PMID: 11006034
-
STAT proteins and transcriptional responses to extracellular signals.
Trends Biochem Sci. 2000 Oct;25(10):496-502
PMID: 11050435
-
Induction of p21WAF1/CIP1 and cyclin D1 expression by the Src oncoprotein in mouse fibroblasts: role of activated STAT3 signaling.
Oncogene. 2000 Nov 16;19(48):5419-27
PMID: 11114718
-
Tumor hypoxia: definitions and current clinical, biologic, and molecular aspects.
J Natl Cancer Inst. 2001 Feb 21;93(4):266-76
PMID: 11181773
-
HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing.
Science. 2001 Apr 20;292(5516):464-8
PMID: 11292862
-
HIF-1, O(2), and the 3 PHDs: how animal cells signal hypoxia to the nucleus.
Cell. 2001 Oct 5;107(1):1-3
PMID: 11595178
-
A conserved family of prolyl-4-hydroxylases that modify HIF.
Science. 2001 Nov 9;294(5545):1337-40
PMID: 11598268
-
Constitutive Stat3 activity up-regulates VEGF expression and tumor angiogenesis.
Oncogene. 2002 Mar 27;21(13):2000-8
PMID: 11960372
-
Constitutive activation of Stat3alpha in brain tumors: localization to tumor endothelial cells and activation by the endothelial tyrosine kinase receptor (VEGFR-2).
Oncogene. 2002 Mar 27;21(13):2058-65
PMID: 11960378
-
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
-
Oxygen sensing and molecular adaptation to hypoxia.
Physiol Rev. 1996 Jul;76(3):839-85
PMID: 8757790
-
STATs and gene regulation.
Science. 1997 Sep 12;277(5332):1630-5
PMID: 9287210
-
Regulation of the hypoxia-inducible transcription factor 1alpha by the ubiquitin-proteasome pathway.
J Biol Chem. 1999 Mar 5;274(10):6519-25
PMID: 10037745
-
STAT3 is a potential modulator of HIF-1-mediated VEGF expression in human renal carcinoma cells.
FASEB J. 2005 Aug;19(10):1296-8
PMID: 15919761
-
Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells.
Exp Mol Med. 2005 Aug 31;37(4):353-64
PMID: 16155412
-
The hypoxia-inducible-factor hydroxylases bring fresh air into hypoxia signalling.
EMBO Rep. 2006 Jan;7(1):41-5
PMID: 16391536
-
STATs in oncogenesis.
Oncogene. 2000 May 15;19(21):2474-88
PMID: 10851046