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Science China Life Sciences

, Volume 57, Issue 5, pp 488–494 | Cite as

Activation of STAT3 stimulates AHSP expression in K562 cells

Open Access
Research Paper
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Abstract

Studies on the chaperone protein α-hemoglobin stabilizing protein (AHSP) reveal that abundant AHSP in erythroid cells enhance the cells’ tolerance to oxidative stress imposed by excess α-hemoglobin in pathological conditions. However, the potential intracellular modulation of AHSP expression itself in response to oxidative stress is still unknown. The present study examined the effect and molecular mechanism of STAT3, an oxidative regulator, on the expression of AHSP. AHSP expression increased in K562 cells upon cytokine IL-6-induced STAT3 activation and decreased in STAT3 knock-down K562 cells. Regulation of AHSP in oxidative circumstance was then examined in α-globin-overloaded K562 cells, and real-time PCR showed strengthened expression of both AHSP and STAT3. ChIP analysis showed binding of STAT3 to AHSP promoter and binding was significantly augmented with IL6 stimulation and upon α-globin overexpression. Dual luciferase reporter assays of the wildtype and mutated SB3 element, an IL-6RE site, in the AHSP promoter in K562 cells highlighted the direct regulatory effect of STAT3 on AHSP gene. Finally, direct binding of STAT3 to SB3 site of AHSP promoter was confirmed with EMSA assays. Our work reveals an adaptive AHSP regulation mediated by the redox-sensitive STAT3 signaling pathway, and provides clues to the therapeutic strategy for AHSP enhancement.

Keywords

AHSP STAT3 oxidative stress IL6-RE 
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References

  1. 1.
    Kihm AJ, Kong YI, Hong W, Russell JE, Rouda S, Adachi K, Simon MC, Blobel GA, Weiss MJ. An abundant erythroid protein that stabilizes free α-haemoglobin. Nature, 2002, 417: 758–763PubMedCrossRefGoogle Scholar
  2. 2.
    Yu X, Kong Y, Dore LC, Abdulmalik O, Katein AM, Zhou S, Choi JK, Gell D, Mackay JP, Gow AJ, Weiss MJ. An erythroid chaperone that facilitates folding of α-globin subunits for hemoglobin synthesis. J Clin Invest, 2007, 117: 1856–1865PubMedPubMedCentralCrossRefGoogle Scholar
  3. 3.
    Feng L, Zhou SP, Gu LC, Gell DA, Mackay JP, Weiss MJ, Gow AJ, Shi Y. Structure of oxidized a-haemoglobin bound to AHSP reveals a protective mechanism for haem. Nature, 2005, 435: 697–701PubMedCrossRefGoogle Scholar
  4. 4.
    Mollan TL, Khandros E, Weiss MJ, Olson JS. Kinetics of α-globin binding to α-hemoglobin stabilizing protein (AHSP) indicate preferential stabilization of hemichrome folding intermediate. J Biol Chem, 2012, 287: 11338–11350PubMedPubMedCentralCrossRefGoogle Scholar
  5. 5.
    Autréaux BD, Toledano MB. ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis. Nat Rev Mol Cell Biol, 2007, 8: 813–824PubMedCrossRefGoogle Scholar
  6. 6.
    Trachootham D, Lu WQ, Ogasawara MA, Nilsa RD, Huang P. Redox regulation of cell survival. Antioxid Redox Signal, 2008, 10: 1343–1374PubMedPubMedCentralCrossRefGoogle Scholar
  7. 7.
    Simon AR, Rai U, Fanburg BL, Cochran BH. Activation of the JAK-STAT pathway by reactive oxygen species. Am J Physiol Cell Physiol, 1998, 275: c1640–c1652Google Scholar
  8. 8.
    Rane SG, Reddy EP. JAKs, STATs and Src Kinases in hematopoiesis. Oncogene, 2002, 21: 3334–3358PubMedCrossRefGoogle Scholar
  9. 9.
    Schindler CW. Series introduction: JAK-STAT signaling in human disease. J Clin Invest, 2002, 109: 1133–1137PubMedPubMedCentralCrossRefGoogle Scholar
  10. 10.
    Menon MP, Karur V, Bogacheva O, Bogachev O, Cuetara B, Wojchowski DM. Signals for stress erythropoiesis are integrated via an erythropoietin receptor-phosphotyrosine-343-Stat5 axis. J Clin Invest, 2006, 116: 683–694PubMedPubMedCentralCrossRefGoogle Scholar
  11. 11.
    Wen ZL, Zhong Z, Darnell JE. Maximal activation of transcription by stat1 and stat3 requires both tyrosine and serine phosphorylation. Cell, 1995, 82: 241–250PubMedCrossRefGoogle Scholar
  12. 12.
    Schuringa JJ, Wierenga AT, Kruijer W, Vellenga E. Consititutive stat3, Tyr705, and Ser727 phosphorylation in acute myeloid leukemia cells caused by the autocrine secretion of interleukin-6. Blood, 2000, 95: 3765–3770PubMedGoogle Scholar
  13. 13.
    Jenkins BJ, Roberts AW, Najdovska M, Grail D, Ernst M. The threshold of gp130-dependent STAT3 signaling is critical for normal regulation of hematopoiesis. Blood, 2005, 105: 3512–3520PubMedCrossRefGoogle Scholar
  14. 14.
    Hou TY, Tieu BC, Ray S, Recinos Iii A, Cui R, Tilton RG, Brasier AR. Roles of IL-6-gp130 signaling in vascular inflammation. Curr Cardiol Rev, 2008, 4: 179–192PubMedPubMedCentralCrossRefGoogle Scholar
  15. 15.
    Wei GH, Zhao GW, Song W, Hao DL, Lv X, Liu DP, Liang CC. Mechanisms of human Γ-globin transcriptional induction by apicidin involves p38 signaling to chromatin. Biochem Biophys Res Commun, 2007, 363: 889–894PubMedCrossRefGoogle Scholar
  16. 16.
    Yao X, Kodeboyina S, Liu L, Dzandu J, Sangerman J, Ofori-Acquah SF, Pace BS. Role of STAT3 and GATA1 interactions in Γ-globin gene expression. Exp hematol, 2009, 37: 889–900PubMedPubMedCentralCrossRefGoogle Scholar
  17. 17.
    Zhou LQ, Wu J, Wang WT, Yu W, Zhao GN, Zhang P, Xiong J, Li M, Xue Z, Wang X, Xie XM, Guo ZC, Lv X, Liu DP. The AT-rich DNA-binding protein SATB2 promotes expression and physical association of human GΓ- and AΓ-globin genes. J Biol Chem, 2012, 287: 30641–30652PubMedPubMedCentralCrossRefGoogle Scholar
  18. 18.
    Wright CJ, Dennery PA. Manipulation of gene expression by oxygen: a primer from bedside to bench. Pediatr Res, 2009, 66: 3–10PubMedPubMedCentralCrossRefGoogle Scholar
  19. 19.
    Darnell JE. STATs and gene regulation. Science, 1997, 277: 1630–1635PubMedCrossRefGoogle Scholar
  20. 20.
    Kojima H, Nakajima K, Hirano T. IL-6-inducible complexes on an IL-6 response element of the junB promoter contain stat3 and 36KD CRE-like site binding protein(s). Oncogene, 1996, 12: 547–554PubMedGoogle Scholar
  21. 21.
    Ichiba M, Nakajima K, Yamanaka Y, Kiuchi N, Hirano T. Autoregulation of the Stat3 gene through cooperation with a cAMP-responsive element-bindding protein. J Biol Chem, 1998, 273: 6132–6138PubMedCrossRefGoogle Scholar
  22. 22.
    Gallagher PG, Liem RI, Wong E, Weiss MJ, Bodine DM. GATA1 and Oct-1 are required for expression of the human α-hemoglobin-stabilizing protein gene. J Biol Chem, 2005, 280: 39016–39023PubMedCrossRefGoogle Scholar
  23. 23.
    Zhao GW, Yang RF, Lü X, Mitchell WJ, Liu DP, Liang CC. NF-E2: a novel regulator of alpha-hemoglobin stabilizing protein gene expression. Chin Med Sci J, 2010, 25: 193–198CrossRefGoogle Scholar
  24. 24.
    Mayani H, Dragowska W, Lansdrop PM. Cytokine-induced selective expansion and maturation of erythroid versus myeloid progenitors from purified core blood precursor cells. Blood, 1993, 81: 3252–3258PubMedGoogle Scholar
  25. 25.
    Gerhartz C, Heesel B, Sasse J, Hemmann U, Landgraf C, Schneider-Mergener J, Horn F, Heinrich PC, Graeve L. Differential activation of acute phase response factor/STAT3 and STAT1 via the cytoplasmic domain of the interleukin 6 signal transducer gp130. J Biol Chem, 1996, 271: 12991–12998PubMedCrossRefGoogle Scholar
  26. 26.
    Watowich SS, Mikami Aki, Busche RA, Xie X, Pharr PN, Longmore GD. Erythropoietin receptors that signal through Stat5 or Stat3 support fetal liver and adult erythropoiesis: lack of specificity of Stat signals during red blood cell development. J Interf Cytok Res, 2000, 20: 1065–1070CrossRefGoogle Scholar
  27. 27.
    Mantel C, Messina-Graham S, Moh A, Cooper S, Hangoc G, Fu XY, Broxmeyer HE. Mouse hematopoietic cell-targeted STAT3 deletion: stem/progenitor cell defects, mitochondrial dysfunction, ROS overproduction, and a rapid aging-like phenotype. Blood, 2012, 120: 2589–2599PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Ingley E, McCarthy DJ, Pore JR, Sarna MK, Adenan AS, Wright MJ, Erber W, Tilbrook PA, Klinken SP. Lyn deficiency reduces GATA-1, EKLF and STAT5, and induces extramedullary stress erythropoiesis. Oncogene, 2005, 24: 336–343PubMedCrossRefGoogle Scholar
  29. 29.
    Röder S, Steimle C, Meinhardt G, Pahl HL. STAT3 is constitutively active in some patients with Polycythemia rubra vera. Exp Hematol, 2001, 29: 694–702PubMedCrossRefGoogle Scholar
  30. 30.
    Dos Santos CO, Duarte ASS, Saad STO, Costa FF. Expression of α-hemoglobin stabilizing protein gene during human erythropoiesis. Exp Hematol, 2004, 32: 157–162PubMedCrossRefGoogle Scholar
  31. 31.
    Wen ZL. Darnell JE. Mapping of Stat3 serine phosphorylation to a single residue (727) and evidence that serine phosphorylation has no influence on DNA binding of Stat1 and Stat3. Nucl Acids Res, 1997, 25: 2062–2067PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Mukaigasa K, Nguyen LTP, Li L, Nakajima H, Yamamoto M, Kobayashi M. Genetic evidence of an evolutionarily conserved role for Nrf2 in the protection against oxidative stress. Mol Cell Biol, 2012, 32: 4455–4461PubMedPubMedCentralCrossRefGoogle Scholar
  33. 33.
    Kawatani Y, Suzuki T, Shimizu R, Kelly VP, Yamamoto M. Nrf2 and selenoproteins are essential for maintaining oxidative homeostasis in erythrocytes and protecting against hemolytic anemia. Blood, 2011, 117: 986–996PubMedCrossRefGoogle Scholar
  34. 34.
    Marro S, Chiabrando D, Messana E, Stolte J, Turco E, Tolosano E, Muckenthaler MU. Heme controls ferroportin1 (FPN1) transcription involving Bach1, Nrf2 and a MARE/ARE sequence motif at position -7007 of the FPN1 promoter. Haematologica, 2010, 95: 1261–1268PubMedPubMedCentralCrossRefGoogle Scholar
  35. 35.
    Macari ER, Lowrey CH. Induction of human fetal hemoglobin via the NRF2 antioxidant response signaling pathway. Blood, 2011, 117: 5987–5997PubMedPubMedCentralCrossRefGoogle Scholar
  36. 36.
    Wruck CJ, Streetz K, Pavic G, Götz ME, Tohidnezhad M, Brandenburg LO, Varoga D, Eickelberg O, Herdegen T, Trautwein C, Cha K, Kan YW, Pufe T. Nrf2 induces interleukin-6(IL-6) expression via antioxidant response element within the IL-6 promoter. J Biol Chem, 2011, 286: 4493–4499PubMedPubMedCentralCrossRefGoogle Scholar
  37. 37.
    Xiong Q, Zhang ZJ, Chang KH, Qu H, Wang H, Qi H, Li Y, Ruan X, Yang Y, Yang Y, Li Y, Sandstrom R, Sabo PJ, Li Q, Stamatoyannopoulos G, Stamatoyannopoulos JA, Fang X. Comprehensive characterization of erythroid-specific enhancers in the genomic regions of human Krüppel-like factors. BMC Genom, 2013, 14: 587CrossRefGoogle Scholar

Copyright information

© The Author(s) 2014

Authors and Affiliations

  1. 1.State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical SciencesChinese Academy of Medical Sciences & Peking Union Medical CollegeBeijingChina

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