Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지

Enhanced Calreticulin Expression Promotes Calcium-dependent Apoptosis in Postnatal Cardiomyocytes

  • Lim, Soyeon (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Chang, Woochul (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Lee, Byoung Kwon (Department of Internal Medicine, Sanggyepaik Hospital, Inje University) ;
  • Song, Heesang (Department of Pediatrics, Washington University in St. Louis School of Medicine) ;
  • Hong, Ja Hyun (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Lee, Sunju (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Song, Byeong-Wook (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Kim, Hye-Jung (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Cha, Min-Ji (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Jang, Yangsoo (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Chung, Namsik (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine) ;
  • Choi, Soon-Yong (Department of Biotechnology, College of Life Science and Nano Technology, Hannam University) ;
  • Hwang, Ki-Chul (Cardiovascular Research Institute, Cardiology Division, Department of Internal Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine)
  • Received : 2007.08.21
  • Accepted : 2008.01.22
  • Published : 2008.05.31
⟨ Previous Next ⟩

Abstract

Calreticulin (CRT) is one of the major $Ca^{2+}$ binding chaperone proteins of the endoplasmic reticulum (ER) and an unusual luminal ER protein. Postnatally elevated expression of CRT leads to impaired development of the cardiac conductive system and may be responsible for the pathology of complete heart block. In this study, the molecular mechanisms that affect $Ca^{2+}$-dependent signal cascades were investigated using CRT-overexpressing cardiomyocytes. In particular, we asked whether calreticulin plays a critical role in the activation of $Ca^{2+}$-dependent apoptosis. In the cells overexpressing CRT, the intracellular calcium concentration was significantly increased and the activity of PKC and level of SECAR2a mRNA were reduced. Phosphorylation of Akt and ERKs decreased compared to control. In addition the activity of the anti-apoptotic factor, Bcl-2, was decreased and the activities of pro-apoptotic factor, Bax, p53 and caspase 8 were increased, leading to a dramatic augmentation of caspase 3 activity. Our results suggest that enhanced CRT expression in mature cardiomyocytes disrupts intracellular calcium regulation, leading to calcium-dependent apoptosis.

Keywords

Acknowledgement

Supported by : Yonsei University College of Medicine, Hannam University

References

  1. Berridge, M.J. (2002). The endoplasmic reticulum: a multifunctional signaling organelle. Cell Calcium 32, 235-249 https://doi.org/10.1016/S0143416002001823
  2. Bonaccorsi, L., Marchiani, S., Muratori, M., Forti, G., and Baldi, E. (2004). Gefitinib ('IRESSA', ZD1839) inhibits EGFinduced invasion in prostate cancer cells by suppressing PI3K/AKT activation. J. Cancer Res. Clin. Oncol. 130, 604-614
  3. Cieslak, D., and Lazou A. (2007). Regulation of BAD protein by PKA, PKCdelta and phosphatases in adult rat cardiac myocytes subjected to oxidative stress. Mol. Cells 24, 224-231
  4. Elia, D., Madhala, D., Ardon, E., Reshef, R., and Halevy, O. (2007). Sonic hedgehog promotes proliferation and differentiation of adult muscle cells: Involvement of MAPK/ERK and PI3K/Akt pathways. Biochim. Biophys. Acta 1773, 1438-1446 https://doi.org/10.1016/j.bbamcr.2007.06.006
  5. Groenendyk, J., Lynch, J., and Michalak, M. (2004). Calreticulin, $Ca^{2+}$, and calcineurin - signaling from the endoplasmic reticulum. Mol. Cells 17, 383-389
  6. Hsieh, Y.C., Jao, H.C., Yang, R.C., Hsu, H.K., and Hsu, C. (2003). Suppression of protein kinase C[alpha] triggers apoptosis through down-regulation of Bcl-xL in a rat hepatic epithelial cell line. Shock 19, 582-587 https://doi.org/10.1097/01.shk.0000065705.84144.ed
  7. Hwang, K.C., Lim, S., Kwon, H.M., Bae, Y.S., Kang, S.M., Chung, K.H., Graham, R.M., Rhee, S.G., and Jang, Y. (2004). Phospholipase C-delta1 rescues intracellular $Ca^{2+}$ overload in ischemic heart and hypoxic neonatal cardiomyocytes. J. Steroid Biochem. Mol. Biol. 91, 131-138 https://doi.org/10.1016/j.jsbmb.2004.02.009
  8. Kageyama, K., Ihara, Y., Goto, S., Urata, Y., Toda, G., Yano, K., and Kondo, T. (2002). Overexpression of caleticulin modulates protein kinase B/Akt signaling to promote apoptosis during cardiac differentiation of cardiomyoblast H9c2 cells. J. Biol. Chem. 277, 19255-19264 https://doi.org/10.1074/jbc.M112377200
  9. Kim, Y.M., Namkoong, S., Yun, Y.G., Hong, H.D., Lee, Y.C., Ha, K.S., Lee, H., Kwon, H.J., Kwon, Y.G., and Kim, Y.M. (2007). Water extract of Korean red ginseng stimulates angiogenesis by activating the PI3K/Akt-dependent ERK1/2 and eNOS pathways in human umbilical vein endothelial cells. Biol. Pharm. Bull. 30, 674-679
  10. Ma, J., and Pan, Z. (2003). Retrograde activation of storeoperated calcium channel. Cell Calcium 33, 375-384 https://doi.org/10.1016/S0143-4160(03)00050-2
  11. Mesaeli, N., Nakamura, K., Zvaritch, E., Dickie, P., Dziak, E., Krause, K.H., Opas, M., MacLennan, D.H., and Michalak, M. (1999). Calreticulin is essential for cardiac development. J. Cell Biol. 144, 857-868 https://doi.org/10.1083/jcb.144.5.857
  12. Michalak, M., Corbett, E.F., Mesaeli, N., Nakamura, K., and Opas, M. (1999). Calreticulin: one protein, one gene, many functions. Biochem. J. 344, 281-292 https://doi.org/10.1042/0264-6021:3440281
  13. Michalak, M., Robert Parker, J.M., and Opas, M. (2002a). $Ca^{2+}$ signaling and calcium binding chaperones of the endoplasmic reticulum. Cell Calcium 32, 269-278 https://doi.org/10.1016/S0143416002001884
  14. Michalak, M., Lynch, J., Groenendyk, J., Guo, L., Parker, J.M.R., and Opas, M. (2002b). Calreticulin in cardiac development and pathology. Biochim. Biophys. Acta 1600, 32-37 https://doi.org/10.1016/S1570-9639(02)00441-7
  15. Milan, D., Griffith, J., Su, M., Price, E.R., and McKeon, F. (1994). The latch region of calcineurin B is involved in both immunosuppressant immunophilin complex docking and phosphatase activation. Cell 79, 437-447 https://doi.org/10.1016/0092-8674(94)90253-4
  16. Moon, D.O., Kim, M.O., Lee, J.D., Choi, Y.H., Lee, M.K., and Kim, G.Y. (2007). Molecular mechanisms of ZD1839 (Iressa)- induced apoptosis in human leukemic U937 cells. Acta Pharmacol. Sin. 28, 1205-1214 https://doi.org/10.1111/j.1745-7254.2007.00615.x
  17. Nakamura, K., Bossy-Wetzel, E., Burns, K., Fadel, M.P., Lozyk, M., Goping, I.S., Opas, M., Bleackley, R.C., Green, D.R., and Michalak, M. (2000). Changes in endoplasmic reticulum luminal environment affect cell sensitivity to apoptosis. J. Cell Biol. 150, 731-740 https://doi.org/10.1083/jcb.150.4.731
  18. Nakamura, K., Robertson, M., Liu, G., Dickie, P., Kyoko, N., Guo, J.Q., Duff, H.J., Opas, M., Kavanaqh, K., and Michalak, M. (2001). Complete heart block and sudden death in mice overexpressing calreticulin. J. Clin. Invest. 107, 1223-1225 https://doi.org/10.1172/JCI13125
  19. Nishimura, K., Blume, P., Ohgi, S., and Sumpio, B.E. (2007). Effect of different frequencies of tensile strain on human dermal fibroblast proliferation and survival. Wound Repair Regen. 5, 646-656
  20. Olson, A.K., Protheroe, K.N., Scholz, T.D., and Segar, J.L. (2008). Activation of the mitogen-activated protein kinases and Akt in response to pulmonary artery banding in the fetal sheep heart is developmentally regulated. Neonatology 93, 145-152 https://doi.org/10.1159/000107729
  21. Papp, S., Dziak, E., Michalak, M., and Opas, M. (2003). Is all of the endoplasmic reticulum created equal? The effects of the heterogeneous distribution of endoplasmic reticulum $Ca^{2+}$- handling proteins. J. Cell Biol. 160, 475-479 https://doi.org/10.1083/jcb.200207136
  22. Periasamy, M., and Huke, S. (2001). SERCA pump level is a critical determinant of $Ca^{2+}$ homeostasis and cardiac contractility. J. Mol. Cell. Cardiol. 33, 1053-1063 https://doi.org/10.1006/jmcc.2001.1366
  23. Pierno, S., Desaphy, J.F., Liantonio, A., De Luca, A., Zarrilli, A., Mastrofrancesco, L., Procino, G., Valenti, G., and Conte Camerino, D. (2007). Disuse of rat muscle in vivo reduces protein kinase C activity controlling the sarcolemma chloride conductance. J. Physiol. 584, 983-995 https://doi.org/10.1113/jphysiol.2007.141358
  24. Pinton, P., Ferrari, D., Rapizzi, E., Virgilio, F.D., Pozzan, T., and Rizzuto, R. (2001). The $Ca^{2+}$ concentration of the endoplasmic reticulum is a key determinant of ceramide-induced apoptosis: significance for the molecular mechanism of Bcl-2 action. EMBO J. 20, 2690-2701 https://doi.org/10.1093/emboj/20.11.2690
  25. Rauch, F., Prud'homme, J., Arabian, A., Dedhar, S., and St- Arnaud, R. (2000). Heart, brain, and body wall defects in mice lacking calreticulin. Exp. Cell Res. 256, 105-111 https://doi.org/10.1006/excr.2000.4818
  26. Salameh, A., Schneider, P., Muhlberg, K., Hagendorff, A., Dhein, S., and Pfeiffer, D. (2004). Chronic regulation of expression of gap junction proteins connexin 40, connexin 43, and connexin 45 in neonatal rat cardiomyocytes. Eur. J. Pharmacol. 503, 9-16 https://doi.org/10.1016/j.ejphar.2004.09.024
  27. Wuytack, F., Raeymaekers, L., and Missiaen, L. (2002). Molecular physiology of the SERCA and SPCA pumps. Cell Calcium 32, 279-305 https://doi.org/10.1016/S0143416002001847