Glucose-regulated protein 75 suppresses apoptosis induced by glucose deprivation in PC12 cells through inhibition of Bax conformational change
Ling Yang
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorXiaoyu Liu
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorJinyu Hao
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorYunlong Yang
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorMingxia Zhao
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorJi Zuo
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorCorresponding Author
Wen Liu
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
*Corresponding author: Tel, 86-21-54237311; Fax, 86-21-54237091; e-mail, [email protected]Search for more papers by this authorLing Yang
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorXiaoyu Liu
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorJinyu Hao
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorYunlong Yang
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorMingxia Zhao
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorJi Zuo
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
Search for more papers by this authorCorresponding Author
Wen Liu
Department of Cellular and Genetic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
*Corresponding author: Tel, 86-21-54237311; Fax, 86-21-54237091; e-mail, [email protected]Search for more papers by this authorThis work was supported by a grant from the Specialized Research Fund for the Doctoral Program of Higher Education of China (No. 20050246079)
Abstract
Glucose-regulated protein 75 (Grp75) is an important molecular chaperone that belongs to the heat shock protein 70 family and resides predominantly in mitochondria. Grp75 can protect cells from glucose deprivation (GD) injury. However, the molecular mechanisms by which it carries out this function are unknown. Here we report that Grp75 could delay the release of cytochrome c and reduce apoptosis induced by GD, and we also found that Grp75 could decrease Bax/Bcl-2 gene expression ratio and inhibit the conformational change of Bax during this process. In conclusion, these findings suggested that Grp75 overexpression was able to inhibit apoptosis induced by GD. Grp75 inhibited Bax conformational change to delay the release of cytochrome c, thus providing protection to PC12 cell which was used primarily as a neuron model against GD toxicity.
References
- 1 Gupta S. Molecular steps of death receptor and mitochondrial pathways of apoptosis. Life Sci 2001, 69: 2957–2964.
- 2 Danial NN, Korsmeyer SJ. Cell death: critical control points. Cell 2004, 116: 205–219.
- 3 Green DR, Kroemer G. The pathophysiology of mitochondrial cell death. Science 2004, 305: 626–629.
- 4 Cory S, Huang DC, Adams JM. The Bcl-2 family: roles in cell survival and oncogenesis. Oncogene 2003, 22: 8590–8607.
- 5 Wall NR, Mohammad RM, Al-Katib AM. Bax: Bcl-2 ratio modulation by bryostatin 1 and novel antitubulin agents is important for susceptibility to drug induced apoptosis in the human early pre-B acute lymphoblastic leukemia cell line, Reh. Leuk Res 1999, 23: 881–888.
- 6 Takahashi Y, Karbowski M, Yamaguchi H, Kazi A, Wu J, Sebti SM, Youle RJ et al. Loss of Bif-1 suppresses Bax/Bak conformational change and mitochondrial apoptosis. Mol Cell Biol 2005, 25: 9369–9382.
- 7 Young JC, Agashe VR, Siegers K, Hartl FU. Pathways of chaperone-mediated protein folding in the cytosol. Nat Rev Mol Cell Biol 2004, 5: 781–791.
- 8 Wadhwa R, Kaul SC, Ikawa Y, Sugimoto Y. Identification of a novel member of mouse hsp70 family. Its association with cellular mortal phenotype. J Biol Chem 1993, 268: 6615–6621.
- 9 Ran Q, Wadhwa R, Kawai R, Kaul SC, Sifers RN, Bick RJ, Smith JR et al. Extramitochondrial localization of mortalin/mthsp70/PBP74/GRP75. Biochem Biophys Res Commun 2000, 275: 174–179.
- 10 Wadhwa R, Taira K, Kaul SC. An Hsp70 family chaperone, mortalin/mthsp70/PBP74/Grp75: what, when, and where Cell Stress Chap-erones 2002, 7: 309–316.
- 11 Liu Y, Liu W, Song XD, Zuo J. Effect of GRP75/mthsp70/PBP74/mortalin overexpression on intracellular ATP level, mitochondrial membrane potential and ROS accumulation following glucose deprivation in PC12 cells. Mol Cell Biochem 2005, 268: 45–51.
- 12 Zheng DH, Zuo J, Yang ZJ, Xia BL, Zhang XN. Grp75 protects cells from injuries caused by glucose deprivation. Yi Chuan Xue Bao 2000, 27: 666–671.
- 13 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-ΔΔCT) method. Methods 2001, 25: 402–408.
- 14
Aubry JP,
Blaecke A,
Lecoanet-Henchoz S,
Jeannin P,
Herbault N,
Caron G,
Moine V
et al.
Annexin V used for measuring apoptosis in the early events of cellular cytotoxicity.
Cytometry
1999, 37: 197–204.
10.1002/(SICI)1097-0320(19991101)37:3<197::AID-CYTO6>3.0.CO;2-L CAS PubMed Web of Science® Google Scholar
- 15 Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ, Roth KA et al. Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 2001, 292: 624–626.
- 16 Hsu YT, Wolter KG, Youle RJ. Cytosol-to-membrane redistribution of Bax and Bcl-X(L) during apoptosis. Proc Natl Acad Sci USA 1997, 94: 3668–3672.
- 17 Wadhwa R, Pereira-Smith OM, Reddel RR, Sugimoto Y, Mitsui Y, Kaul SC. Correlation between complementation group for immortality and the cellular distribution of mortalin. Exp Cell Res 1995, 216: 101–106.
- 18 Kaula SC, Reddelb RR, Sugiharac T, Mitsuia Y, Wadhwac R. Inacti-vation of p53 and life span extension of human diploid fibroblasts by mot-2. FEBS Lett 2000, 474: 159–164.
- 19 Xu J, Xiao HH, Sartorelli AC. Attenuation of the induced differentiation of HL-60 leukemia cells by mitochondrial chaperone HSP70. Oncol Res 1999, 11: 429–435.
- 20 Mizukoshi E, Suzuki M, Loupatov A, Uruno T, Hayashi H, Misono T, Kaul SC et al. Fibroblast growth factor-1 interacts with the glucose-regulated protein GRP75/mortalin. Biochem J 1999, 343: 461–466.
- 21 Taurin S, Seyrantepe V, Orlov SN, Tremblay TL, Thibault P, Bennett MR, Hamet P et al. Proteome analysis and functional expression identify mortalin as an antiapoptotic gene induced by elevation of [Na+]i/[K+]i ratio in cultured vascular smooth muscle cells. Circ Res 2002, 91: 915–922.
- 22 Massa SM, Longo FM, Zuo J, Wang S, Chen J, Sharp FR. Cloning of rat grp75, an hsp70-family member, and its expression in normal and ischemic brain. J Neurosci Res 1995, 40: 807–819.
- 23 Reed JC, Green DR. Remodeling for demolition: Changes in mitochondrial ultrastructure during apoptosis. Mol Cell 2002, 9: 1–3.
- 24 Kulms D, Schwarz T. Molecular mechanisms involved in UV-induced apoptotic cell death. Skin Pharmacol Appl Skin Physiol 2002, 15: 342–347.
- 25 Goyeneche AA, Harmon JM, Telleria CM. Cell death induced by serum deprivation in luteal cells involves the intrinsic pathway of apoptosis. Reproduction 2006, 131: 103–111.
- 26 Farrow SN, Brown R. New members of the Bcl-2 family and their protein partners. Curr Opin Genet Dev 1996, 6: 45–49.
- 27 Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI et al. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 1997, 275: 1129–1132.
- 28 Zha H, Reed JC. Heterodimerization-independent functions of cell death regulatory proteins Bax and Bcl-2 in yeast and mammalian cells. J Biol Chem 1997, 272: 31482–31488.
- 29 Schinzel A, Kaufmann T, Schuler M, Martinalbo J, Grubb D, Borner C. Conformational control of Bax localization and apoptotic activity by Pro168. J Cell Biol 2004, 164: 1021–1032.
- 30 Samali A, Cotter TG. Heat shock proteins increase resistance to apoptosis. Exp Cell Res 1996, 223: 163–170.
- 31 McClellan AJ, Frydman J. Molecular chaperones and the art of recognizing a lost cause. Nat Cell Biol 2001, 3: E51–E53.
- 32 Hartl FU, Hayer-Hartl M. Molecular chaperones in the cytosol: from nascent chain to folded protein. Science 2002, 295: 1852–1858.
- 33 Wadhwa R, Yaguchi T, Hasan MK, Mitsui Y, Reddel RR, Kaul SC. Hsp70 family member, mot-2/mthsp70/GRP75, binds to the cyto-plasmic sequestration domain of the p53 protein. Exp Cell Res 2002, 274: 246–253.
- 34 Chipuk JE, Kuwana T, Bouchier-Hayes L, Droin NM, Newmeyer DD, Schuler M, Green DR. Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science 2004, 303: 1010–1014.