Abstract
Aim:
To investigate the mechanisms underlying the biphasic redox regulation of hypoxia-inducible factor-1 (HIF-1) transcriptional activity under different levels of oxidative stress caused by reactive oxidative species (ROS).
Methods:
HeLa cells were exposed to different concentrations of H2O2 as a simple model for mild and severe oxidative stress. Luciferase reporter assay and/or quantitative real-time PCR were used to investigate the transcriptional activity. Immunoblot was used to detect protein expression. Chromatin immunoprecipitation assay was used to detect HIF-1/DNA binding. The interaction of p300 with HIF-1α or with SENP3, and the SUMO2/3 conjugation states of p300 were examined by coimmunoprecipitation.
Results:
HIF-1 transcriptional activity in HeLa cells was enhanced by low doses (0.05–0.5 mmol/L) of H2O2, but suppressed by high doses (0.75–8.0 mmol/L) of H2O2. The amount of co-activator p300 bound to HIF-1α in HeLa cells was increased under mild oxidative stress, but decreased under severe oxidative stress. The ROS levels differentially modified cysteines 243 and 532 in the cysteine protease SENP3, regulating the interaction of SENP3 with p300 to cause different SUMOylation of p300, thus shifting HIF-1 transcriptional activity.
Conclusion:
The shift of HIF-1 transactivation by ROS is correlated with and dependent on the biphasic redox sensing of SENP3 that leads to the differential SENP3/p300 interaction and the consequent fluctuation in the p300 SUMOylation status.
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Finkel T . Signal transduction by reactive oxygen species. J Cell Biol 2011; 194: 7–15.
Ramsey MR, Sharpless NE . ROS as a tumour suppressor? Nat Cell Biol 2006; 8: 1213–15.
Schulz TJ, Zarse K, Voigt A, Urban N, Birringer M, Ristow M . Glucose restriction extends Caenorhabditis elegans life span by inducing mitochondrial respiration and increasing oxidative stress. Cell Metab 2007; 6: 280–93.
Gough DR, Cotter TG . Hydrogen peroxide: a Jekyll and Hyde signalling molecule. Cell Death Dis 2011; 2: e213. doi: 10.1038/cddis.96.
Mesquita A, Weinberger M, Silva A, Sampaio-Marques B, Almeida B, Leão C, et al. Caloric restriction or catalase inactivation extends yeast chronological lifespan by inducing H2O2 and superoxide dismutase activity. Proc Natl Acad Sci U S A 2010; 107: 15123–8.
Martindale JL, Holbrook NJ . Cellular response to oxidative stress: signaling for suicide and survival. J Cell Physio 2002; 192: 1–15.
Gao L, Mann GE . Vascular NAD(P)H oxidase activation in diabetes: a double-edged sword in redox signalling. Cardiovasc Res 2009; 82: 9–20.
Ray PD, Huang BW, Tsuji Y . Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal 2012; 24: 981–90.
Cramer T, Yamnishi Y, Clausen BE, Förster I, Pawlinski R, Mackman N, et al. HIF-1alpha is essential for myeloid cell-mediated inflammation. Cell 2003; 112: 645–57.
Jung YJ, Isaacs JS, Lee S, Trepel J, Neckers L . IL-1beta-mediated up-regulation of HIF-1alpha via an NFkappaB/COX-2 pathway identifies HIF-1 as a critical link between inflammation and oncogenesis. FASEB J 2003; 17: 2115–7.
Walmsley SR, Cadwallader KA, Chilvers ER . The role of HIF-1alpha in myeloid cell inflammation. Trends Immunol 2005; 26: 434–9.
Tacchini L, Dansi P, Matteucci E, Desiderio MA . Hepatocyte growth factor signalling stimulates hypoxia inducible factor-1 (HIF-1) activity in HepG2 hepatoma cells. Carcinogenesis 2001; 22: 1363–71.
Patten DA, Lafleur VN, Robitaille GA, Chan DA, Giaccia AJ, Richard DE . Hypoxia-inducible factor-1 activation in nonhypoxic conditions: the essential role of mitochondrial-derived reactive oxygen species. Mol Biol Cell 2010; 21: 3247–57.
Zelzer E, Levy Y, Kahana C, Shilo BZ, Rubinstein M, Cohen B . Insulin induces transcription of target genes through the hypoxia-inducible factor HIF-1alpha/ARNT. EMBO J 1998; 17: 5085–94.
Carroll VA, Ashcroft M . Role of hypoxia-inducible factor (HIF)-1alpha versus HIF-2alpha in the regulation of HIF target genes in response to hypoxia, insulin-like growth factor-I, or loss of von Hippel-Lindau function: implications for targeting the HIF pathway. Cancer Res 2006, 66: 6264–70.
Roth U, Curth K, Unterman TG, Kietzmann T . The transcription factors HIF-1 and HNF-4 and the coactivator p300 are involved in insulin-regulated glucokinase gene expression via the phosphatidylinositol 3-kinase/protein kinase B pathway. J Biol Chem 2004; 279: 2623–31.
Treins C, Murdaca J, Van Obberghen E, Giorgetti-Peraldi S . AMPK activation inhibits the expression of HIF-1alpha induced by insulin and IGF-1. Biochem Biophys Res Commun 2006; 342: 1197–202.
Chandel NS, Maltepe E, Goldwasser E, Mathieu CE, Simon MC, Schumacker PT . Mitochondrial reactive oxygen species trigger hypoxia-induced transcription. Proc Natl Acad Sci U S A 1998; 95: 11715–20.
Chandel NS, Budinger G . The cellular basis for diverse responses to oxygen. Free Radic Biol Med 2007; 42: 165–74.
Jung SN, Yang W, Kim J, Kim HS, Kim EJ, Yun H, et al. 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 2008; 29: 713–21.
Huang XZ, Wang J, Huang C, Chen YY, Shi GY, Hu QS, et al. Emodin enhances cytotoxicity of chemotherapeutic drugs in prostate cancer cells: the mechanisms involve ROS-mediated suppression of multidrug resistance and hypoxia inducible factor-1. Cancer Biol Ther 2008; 7: 468–75.
Galanis A, Pappa A, Giannakakis A, Lanitis E, Dangaj D, Sandaltzopoulos R . Reactive oxygen species and HIF-1 signalling in cancer. Cancer Lett 2008; 266: 12–20.
Tuttle SW, Maity A, Oprysko PR, Kachur AV, Ayene IS, Biaglow JE, et al. Detection of reactive oxygen species via endogenous oxidative pentose phosphate cycle activity in response to oxygen concentration: implications for the mechanism of HIF-1alpha stabilization under moderate hypoxia. J Biol Chem 2007; 282: 36790–6.
Comito G, Calvani M, Giannoni E, Bianchini F, Calorini L, Torre E, et al. HIF-1α stabilization by mitochondrial ROS promotes Met-dependent invasive growth and vasculogenic mimicry in melanoma cells. Free Radic Biol Med 2011; 51: 893–904.
Schumacker PT . SIRT3 controls cancer metabolic reprogramming by regulating ROS and HIF. Cancer Cell 2011; 19: 299–300.
Huang C, Han Y, Wang YM, Sun XX, Yan S, Yeh ET, et al. SENP3 is responsible for HIF-1 transactivation under mild oxidative stress via p300 de-SUMOylation. EMBO J 2009; 28: 2748–62.
Han Y, Huang C, Sun X, Xiang B, Wang M, Yeh ET, et al. SENP3-mediated de-conjugation of SUMO2/3 from promyelocytic leukemia is correlated with accelerated cell proliferation under mild oxidative stress. J Biol Chem 2010; 285: 12906–15.
Xu Z, Lam LS, Lam LH, Chau SF, Ng TB, Au SW . Molecular basis of the redox regulation of SUMO proteases. FASEB J 2008; 22: 127–37.
Gong L, Yeh ET . Characterization of a family of nucleolar SUMO-specific proteases with preference for SUMO-2 or SUMO-3. J Biol Chem 2006; 281: 15869–77.
Wang Y, Mukhopadhyay D, Mathew S, Hasebe T, Heimeier RA, Azuma Y, et al. Identification and developmental expression of Xenopus laevis SUMO proteases. PLoS One 2009; 4: e8462.
Bossis G, Melchior F . Regulation of SUMOylation by reversible oxidation of SUMO conjugating enzymes. Mol Cell 2006; 21: 349–57.
Yi J, Yang J, He R, Gao F, Sang H, Tang XM, et al. Emodin enhances arsenic trioxide induced apoptosis via generation of reactive oxygen species and inhibition of survival signaling. Cancer Res 2004; 64: 108–16.
Zuo Y, Xiang BG, Yang J, Sun XX, Wang YM, Cang H, et al. oxidative modification of caspase-9 facilitates its activation via disulfide-mediated interaction with Apaf-1. Cell Res 2009; 19: 449–57.
Sang J, Yang K, Sun YP, Han Y, Cang H, Chen YY, et al. SUMO2 and SUMO3 transcription is differentially regulated by oxidative stress in an Sp1-dependent manner. Biochem J 2011; 435: 489–98.
Yan S, Sun XX, Xiang BG, Cang H, Kang XL, Chen YY, et al. Redox regulation of the stability of the SUMO protease SENP3 via interactions with CHIP and Hsp90. EMBO J 2010; 29: 3773–86.
Lando D, Peet D, Whelan DA, Gorman JJ, Whitelaw ML . Asparagine hydroxylation of the HIF transactivation domain a hypoxic switch. Science 2002; 295: 858–61.
Kallio PJ, Okamoto K, O'Brien S, Carrero P, Makino Y, Tanaka H, et al. Signal transduction in hypoxic cells: inducible nuclear translocation and recruitment of the CBP/p300 coactivator by the hypoxia-inducible factor-1alpha. EMBO J 1998; 17: 6573–86.
Freedman SJ, Sun ZY, Poy F, Kung AL, Livingston DM, Wagner G, et al. Structural basis for recruitment of CBP/p300 by hypoxia-inducible factor-1 alpha. Proc Natl Acad Sci U S A 2002; 99: 5367–72.
Ebert BL, Bunn HF . Regulation of transcription by hypoxia requires a multiprotein complex that includes hypoxia-inducible factor 1, an adjacent transcription factor, and p300/CREB binding protein. Mol Cell Biol 1998; 18: 4089–96.
Huang LE, Arany Z, Livingston DM, Bunn HF . Activation of hypoxia-inducible transcription factor depends primarily upon redox-sensitive stabilization of its alpha subunit. J Biol Chem 1996; 271: 32253–9.
Irani K . Oxidant signaling in vascular cell growth, death, and survival: a review of the roles of reactive oxygen species in smooth muscle and endothelial cell mitogenic and apoptotic signaling. Circ Res 2000; 87: 179–83.
Deshpande SS, Angkeow P, Huang J, Ozaki M, Irani K . Rac1 inhibits TNF-alpha-induced endothelial cell apoptosis: dual regulation by reactive oxygen species. FASEB J 2000; 14: 1705–14.
Lopes N, Gregg D, Vasudevan S, Hassanain H, Goldschmidt-Clermont P, Kovacic H . Thrombospondin 2 regulates cell proliferation induced by Rac1 redox-dependent signaling. Mol Cell Biol 2003; 23: 5401–8.
Day RM, Suzuki YJ . Cell proliferation, reactive oxygen and cellular glutathione. Dose Response 2006; 3: 425–42.
Jing YW, Yang J, Wang YM, Li H, Chen YY, Hu QS, et al. Alteration of subcellular redox equilibrium and the consequent oxidative modification of nuclear factor kappaB are critical for anticancer cytotoxicity by emodin, a reactive oxygen species-producing agent. Free Radic Biol Med 2006; 40: 2183–97.
Wang YM, Huang XZ, Cang H, Gao F, Yamamoto T, Osaki T, et al. The endogenous reactive oxygen species promote NF-kappaB activation by targeting on activation of NF-kappaB-inducing kinase in oral squamous carcinoma cells. Free Radic Res 2007; 41: 963–71.
Yi J, Yang J, He R, Gao F, Sang H, Tang XM, et al. Emodin enhances arsenic trioxide-induced apoptosis via generation of reactive oxygen species and inhibition of survival signaling. Cancer Res 2004; 64: 108–16.
Wang J, Yi J . Cancer cell killing via ROS: to increase or decrease, that is the question. Cancer Biol Ther 2008; 7: 1875–84.
Thamsen M, Jakob U . The redoxome: proteomic analysis of cellular redox networks. Curr Opin Chem Biol 2011; 15: 113–9.
Winter J, Linke K, Jatzek A, Jakob U . Severe oxidative stress causes inactivation of DnaK and activation of the redox-regulated chaperone Hsp33. Mol Cell 2005; 17: 381–92.
Leichert LI, Gehrke F, Gudiseva HV, Blackwell T, Ilbert M, Walker AK, et al. Quantifying changes in the thiol redox proteome upon oxidative stress in vivo. Proc Natl Acad Sci U S A 2008; 105: 8197–202.
Biswas S, Chida AS, Rahman I . Redox modifications of protein-thiols: emerging roles in cell signaling. Biochem Pharmacol 2006; 71: 551–64.
Eaton P . Protein thiol oxidation in health and disease: techniques for measuring disulfides and related modifications in complex protein mixtures. Free Radic Biol Med 2006; 40: 1889–99.
Wang Y, Yang J, Yi J . Redox sensing by proteins: oxidative modifications on cysteines and the consequent events. Antioxid Redox Signal 2012; 16: 649–57.
Liu XH, Kirschenbaum A, Lu M, Yao S, Dosoretz A, Holland JF, et al. Prostaglandin E 2 induces hypoxia-inducible factor-1alpha stabilization and nuclear localization in a human prostate cancer cell line. J Biol Chem 2002; 277: 50081–6.
Sun W, Chang SS, Fu Y, Liu Y, Califano JA . Chronic CSE treatment induces the growth of normal oral keratinocytes via PDK2 upregulation, increased glycolysis and HIF1α stabilization. PLoS One 2011; 6: e16207.
Mao Y, Song G, Cai Q, Liu M, Luo H, Shi M, et al. Hydrogen peroxide-induced apoptosis in human gastric carcinoma MGC803 cells. Cell Biol Int 2006; 30: 332–7.
Hardwick JS, Sefton BM . Activation of the Lck tyrosine protein kinase by hydrogen peroxide requires the phosphorylation of Tyr-394. Proc Natl Acad Sci U S A 1995; 92: 4527–31.
Sullivan SG, Chiu DT, Errasfa M, Wang JM, Qi JS, Stern A . Effects of H2O2 on protein tyrosine phosphatase activity in HER14 cells. Free Radic Biol Med 1994; 16: 399–403.
Chou WC, Dang CV . Acute promyelocytic leukemia: recent advances in therapy and molecular basis of response to arsenic therapies. Curr Opin Hematol 2005; 12: 1–6.
Lallemand-Breitenbach V, Jeanne M, Benhenda S, Nasr R, Lei M, Peres L, et al. Arsenic degrades PML or PML-RARalpha through a SUMO-triggered RNF4/ubiquitin-mediated pathway. Nat Cell Biol 2008; 10: 547–55.
Acknowledgements
This study was supported by grants from the National Natural Science Foundation of China (91013012, 30971437 to Jing YI, 31000613 to Xin-zhi HUANG), Shanghai Municipal Science and Technology Commission (11ZR1419100 to Ying WANG, 11DZ2260200 to Jing YI), and Shanghai Municipal Education Commission (J50201, to Jing YI).
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary figures are available at website of Acta Pharmacologica Sinica on NPG.
Supplementary information
Supplementary information, Figure S1
The procedures were as the same as figue 1C. (DOC 52 kb)
Supplementary information, Figure S2
HeLa cells were transfected with RGS-SENP3 for 48 h. (DOC 84 kb)
Rights and permissions
About this article
Cite this article
Wang, Y., Yang, J., Yang, K. et al. The biphasic redox sensing of SENP3 accounts for the HIF-1 transcriptional activity shift by oxidative stress. Acta Pharmacol Sin 33, 953–963 (2012). https://doi.org/10.1038/aps.2012.40
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/aps.2012.40
Keywords
This article is cited by
-
SUMO-specific protease 3 is a key regulator for hepatic lipid metabolism in non-alcoholic fatty liver disease
Scientific Reports (2016)
-
Hydrogen peroxide – production, fate and role in redox signaling of tumor cells
Cell Communication and Signaling (2015)
-
SUMO-specific proteases/isopeptidases: SENPs and beyond
Genome Biology (2014)
-
P300-dependent STAT3 acetylation is necessary for angiotensin II-induced pro-fibrotic responses in renal tubular epithelial cells
Acta Pharmacologica Sinica (2014)
