Abstract
We previously report that BMP2 mediates mild ER stress-activated ATF6 and directly regulates XBP1S splicing in the course of chondrogenesis. The mammalian unfolded protein response (UPR) protects the cell against the stress of misfolded proteins in the endoplasmic reticulum (ER). Failure to adapt to ER stress causes the UPR to trigger apoptosis. The transcription factor activating transcription factor 6 (ATF6), a key regulator of the UPR, is known to be important for ER stress-mediated apoptosis and cell growth, but the molecular mechanism underlying these processes remains unexplored. In this study, we demonstrate that ATF6 is differentially expressed during BMP2-stimulated chondrocyte differentiation and exhibits prominent expression in growth plate chondrocytes. ATF6 can enhance the level of IRE1a-spliced XBP1S protein in chondrogenesis. IRE1a and ATF6 can synergistically regulate endogenous XBP1S gene expression in chondrogenesis. Furthermore, overexpression ATF6 inhibited, while ATF6-knockdown enhanced, the cell proliferation in chondrocyte development with G1 phase arresting, S phase reducing and G2-M phase delaying. Besides, Ad-ATF6 can activate, whereas knockdown ATF6 by an siRNA-silencing approach inhibited, ER stress-mediated apoptosis in chondrogenesis induced by BMP2, as assayed by cleaved caspase3, CHOP, p-JNK expression in the course of chondrocyte differentiation. On the other hand, FCM, TUNEL assay and immunohistochemistry analysis also proved this result in vitro and in vivo. It was demonstrated that Ad-ATF6 activation of the ER stress-specific caspase cascade in developing chondrocyte tissue. Collectively, these findings reveal a novel critical role of ATF6 in regulating ER stress-mediated apoptosis in chondrocyte differentiation and the molecular mechanisms involved.
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10 August 2020
The Editor-in-Chief has retracted the article by Han et al. (2014) because Fig. 3a-d are also published as Fig. 5b-e in Liu et al. (2012), and Fig. 3a, c, d are also published as Fig. 5a, d, e in Guo et al. (2014).
References
Back SH, Lee K, Vink E, Kaufman RJ (2006) Cytoplasmic IRE1alpha-mediated XBP1 mRNA splicing in the absence of nuclear processing and endoplasmic reticulum stress. J Biol Chem 281(27):18691–18706
Bernales S, Papa FR, Walter P (2006) Intracellular signaling by the unfolded protein response. Annu Rev Cell Dev Biol 22:487–508
Canalis E, Economides AN, Gazzerro E (2003) Bone morphogenetic proteins, their antagonists, and the skeleton. Endocr Rev 24(2):218–235
Degterev A, Boyce M, Yuan J (2003) A decade of caspases. Oncogene 22(53):8543–8567
Fulda S, Gorman AM, Hori O, Samali A (2010) Cellular stress responses: cell survival and cell death. Int J Cell Biol 2010:214074
Furuichi T, Ikegawa S, Ikawa M, Okabe M, Wanaka A, Imaizumi K (2009) Signalling mediated by the endoplasmic reticulum stress transducer OASIS is involved in bone formation. Nat Cell Biol 11:1205–1211
Gotoh T, Oyadomari S, Mori K, Mori M (2002) Nitric oxide-induced apoptosis in RAW 264.7 macrophages is mediated by endoplasmic reticulum stress pathway involving ATF6 and CHOP. J Biol Chem 277(14):12343–12350
Guo F, Lin EA, Liu P, Lin J, Liu C (2010) XBP1U inhibits the XBP1S-mediated upregulation of the iNOS gene expression in mammalian ER stress response. Cell Signal 22(12):1818–1828
Guo FJ, Liu Y, Zhou J, Luo S, Zhao W, Li X, Liu C (2012) XBP1S protects cells from ER stress-induced apoptosis through Erk1/2 signaling pathway involving CHOP. Histochem Cell Biol 138(3):447–460
Hassler J, Cao SS, Kaufman RJ (2012) IRE1, a double-edged sword in pre-miRNA slicing and cell death. Dev Cell 23(5):921–923
Hitomi J, Katayama T, Eguchi Y, Kudo T, Taniguchi M, Koyama Y, Manabe T, Yamagishi S, Bando Y, Imaizumi K, Tsujimoto Y, Tohyama M (2004) Involvement of caspase-4 in endoplasmic reticulum stress-induced apoptosis and Abeta-induced cell death. J Cell Biol 165(3):347–356
Iwakoshi NN, Lee AH, Vallabhajosyula P, Otipoby KL, Rajewsky K, Glimcher LH (2003) Plasma cell differentiation and the unfolded protein response intersect at the transcription factor XBP-1. Nat Immunol 4:321–329
Jang WG, Kim EJ, Kim DK, Ryoo HM, Lee KB, Kim SH, Choi HS, Koh JT (2012) BMP2 protein regulates osteocalcin expression via Runx2-mediated ATF6 gene transcription. J Biol Chem 287:905–915
Kaufman RJ (2002) Orchestrating the unfolded protein response in health and disease. J Clin Invest 110(10):1389–1398
Kim R, Emi M, Tanabe K, Murakami S (2006) Role of the unfolded protein response in cell death. Apoptosis 11(1):5–13
Lefebvre V, Smits P (2005) Transcriptional control of chondrocyte fate and differentiation. Birth Defects Res C Embryo Today 75(3):200–212
Li M, Baumeister P, Roy B, Phan T, Foti D, Luo S, Lee AS (2000) ATF6 as a Transcription Activator of the endoplasmic reticulum stress element: thapsigargin stress-induced changes and synergistic interactions with NF-Y and YY1. Mol Cell Biol USA 20(14):5096–5106
Liu CY, Schröder M, Kaufman RJ (2000) Ligand-independent dimerization activates the stress response kinases IRE1 and PERK in the lumen of the endoplasmic reticulum. J Biol Chem 275(32):24881–24885
Liu CY, Xu Z, Kaufman RJ (2003) Structure and intermolecular interactions of the luminal dimerization domain of human IRE1alpha. J Biol Chem 278(20):17680–17687
Liu Y, Zhou J, Zhao W, Li X, Jiang R, Liu C, Guo FJ (2012) XBP1S associates with RUNX2 and regulates chondrocyte hypertrophy. J Biol Chem 287(41):34500–34513
Luan Y, Yu XP, Xu K, Ding B, Yu J, Huang Y, Yang N, Lengyel P, Di Cesare PE, Liu CJ (2007) The retinoblastoma protein is an essential mediator of osteogenesis that links the p204 protein to the Cbfa1 transcription factor thereby increasing its activity. J Biol Chem 282:16860–16870
Luan Y, Yu XP, Yang N, Frenkel S, Chen L, Liu CJ (2008) p204 protein overcomes the inhibition of Cbfa1-mediated Osteogenic differentiation by Id helix-loop-helix proteins. Mol Biol Cell 19:2113–2126
Lui JC, Andrade AC, Forcinito P, Hegde A, Chen W, Baron J, Nilsson O (2010) Spatial and temporal regulation of gene expression in the mammalian growth plate. Bone 46(5):1380–1390
Morishima N, Nakanishi K, Takenouchi H, Shibata T, Yasuhiko Y (2002) An ER stress-specific caspase cascade in apoptosis: cytochrome c-independent activation of caspase-9 by caspase-12. J Biol Chem 277(37):34287–34294
Murakami T, Saito A, Hino S, Kondo S, Kanemoto S, Chihara K, Sekiya H, Tsumagari K, Ochiai K, Yoshinaga K, Saitoh M, Nishimura R, Yoneda T, Kou I, Jang WG, Kim EJ, Kim DK, Ryoo HM, Lee KB, Kim SH, Choi HS, Koh JT (2012) BMP2 protein regulates osteocalcin expression via Runx2-mediated ATF6 gene transcription. J Biol Chem 287(2):905–915
Nakagawa T, Yuan J (2000) Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis. J Cell Biol 150:804–887
Nakagawa T, Zhu H, Morishima N, Li E, Xu J, Yankner BA, Yuan J (2000) Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta. Nature 403(6765):98–103
Nakanishi Keiko, Sudo Tatsuhiko, Morishima Nobuhiro (2005) Endoplasmic reticulum stress signaling transmitted by ATF6 mediates apoptosis during muscle development. J Cell Biol 169(4):555–560
Ron D, Walter P (2007) Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 8(7):519–529
Saito A, Ochiai K, Kondo S, Tsumagari K, Murakami T, Cavener DR, Imaizumi K (2011) Endoplasmic reticulum stress response mediated by the PERK-eIF2(alpha)-ATF4 pathway is involved in osteoblast differentiation induced by BMP2. J Biol Chem 286(6):4809–4818
Samali A, Zhivotovsky B, Jones D, Nagata S, Orrenius S (1999) Apoptosis: cell death defined by caspase activation. Cell Death Differ 6(6):495–496
Welch RD, Jones AL, Bucholz RW, Reinert CM, Tjia JS, Pierce WA, Wozney JM, Li XJ (1998) Effect of recombinant human bone morphogenetic protein-2 on fracture healing in a goat tibial fracture model. J Bone Miner Res 13(9):1483–1490
Zuscik Michael J, Hilton Matthew J, Zhang Xinping, Chen Di, O’Keefe Regis J (2008) Regulation of chondrogenesis and chondrocyte differentiation by stress. J Clin Invest 118:429–438
Acknowledgments
This work was supported by the National Science Foundation of China (No. 81371928, No. 81171697); New Century Excellent Talent Support Project of Education Ministry of China (NCET-12-1090); Returned Overseas Preferred Funded Project of Human Resources and Social Security Ministry of China (No. 2011-235).
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This article has been retracted. Please see the retraction notice for more detail:https://doi.org/10.1007/s00418-020-01907-4"
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Han, X., Zhang, P., Jiang, R. et al. RETRACTED ARTICLE: Explore on the effect of ATF6 on cell growth and apoptosis in cartilage development. Histochem Cell Biol 142, 497–509 (2014). https://doi.org/10.1007/s00418-014-1233-y
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DOI: https://doi.org/10.1007/s00418-014-1233-y