Biochemistry (Moscow)

, Volume 78, Issue 1, pp 91–95 | Cite as

Localization of non-native D-glyceraldehyde-3-phosphate dehydrogenase in growing and apoptotic HeLa cells

  • E. I. Arutyunova
  • L. V. Domnina
  • A. A. Chudinova
  • O. N. Makshakova
  • D. Y. Arutyunov
  • V. I. MuronetzEmail author


Monoclonal antibodies that could not bind native tetramers of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) but could bind to dimeric, monomeric, or denatured forms of GAPDH were used to investigate its intracellular localization. These antibodies distinctly stained the nucleus in growing HeLa cells. In the cytoplasm, non-native GAPDH was colocalized with actin filaments. Incubation of HeLa cells with tumor necrosis factor α (TNF-α) and the protein synthesis inhibitor emetine led to a drastic increase in the amount of the non-native GAPDH in the nuclei. Overproduction of Bcl-2 protein did not change the non-native GAPDH localization in the growing HeLa cells but prevented the development of apoptosis and the increase in the amount of non-native GAPDH in the nuclei upon incubation with TNF-α.

Key words

GAPDH actin HeLa localization apoptosis TNF-α 



fluorescein isothiocyanate


phosphorylating D-glyceraldehyde-3-phosphate dehydrogenase (EC


monoclonal antibodies


tumor necrosis factor α


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  1. 1.
    Nagradova, N. K. (2001) Biochemistry (Moscow), 66, 1067–1076.CrossRefGoogle Scholar
  2. 2.
    Bryksin, A. V., and Laktionov, P. P. (2008) Biochemistry (Moscow), 73, 619–625.CrossRefGoogle Scholar
  3. 3.
    Sirover, M. A. (2011) Biochim. Biophys. Acta, 1810, 741–751.PubMedCrossRefGoogle Scholar
  4. 4.
    Sundararaj, K. P., Wood, R. E., Ponnusamy, S., Salas, A. M., Szulc, Z., Bielawska, A., Obeid, L. M., Hannun, Y. A., and Ogretmen, B. (2004) J. Biol. Chem., 279, 6152–6162.PubMedCrossRefGoogle Scholar
  5. 5.
    Meyer-Siegler, K., Mauro, D. J., Seal, G., Wurzer, J., deRiel, J. K., and Sirover, M. A. (1991) Proc. Natl. Acad. Sci. USA, 88, 8460–8464.PubMedCrossRefGoogle Scholar
  6. 6.
    Azam, S., Jouvet, N., Jilani, A., Vongsamphanh, R., Yang, X., Yang, S., and Ramotar, D. (2008) J. Biol. Chem., 283, 30632–30641.PubMedCrossRefGoogle Scholar
  7. 7.
    Zheng, L., Roeder, R. G., and Luo, Y. (2003) Cell, 114, 255–266.PubMedCrossRefGoogle Scholar
  8. 8.
    Kim, S., Lee, J., and Kim, J. (2007) Biochem. J., 404, 197–206.PubMedCrossRefGoogle Scholar
  9. 9.
    Carujo, S., Estanyol, J. M., Ejarque, A., Agell, N., Bachs, O., and Pujol, M. J. (2006) Oncogene, 25, 4033–4042.PubMedCrossRefGoogle Scholar
  10. 10.
    Saunders, P. A., Chen, R. W., and Chuang, D. M. (1999) J. Neurochem., 72, 925–932.PubMedCrossRefGoogle Scholar
  11. 11.
    Brown, V. M., Krynetski, E. Y., Krynetskaia, N. F., Grieger, D., Mukatira, S. T., Murti, K. G., Slaughter, C. A., Park, H. W., and Evans, W. E. (2004) J. Biol. Chem., 279, 5984–5992.PubMedCrossRefGoogle Scholar
  12. 12.
    Park, J., Han, D., Kim, K., Kang, Y., and Kim, Y. (2009) Biochim. Biophys. Acta, 1794, 254–262.PubMedCrossRefGoogle Scholar
  13. 13.
    Kornberg, M. D., Sen, N., Hara, M. R., Juluri, K. R., Nguyen, J. V., Snowman, A. M., Law, L., Hester, L. D., and Snyder, S. H. (2010) Nat. Cell Biol., 12, 1094–1100.PubMedCrossRefGoogle Scholar
  14. 14.
    Ventura, M., Mateo, F., Serratosa, J., Salaet, I., Carujo, S., Bachs, O., and Pujol, M. J. (2010) Int. J. Biochem. Cell. Biol., 42, 1672–1680.PubMedCrossRefGoogle Scholar
  15. 15.
    Schmitz, H. D. (2001) Eur. J. Cell Biol., 80, 419–427.PubMedCrossRefGoogle Scholar
  16. 16.
    Fokina, K. V., Dainyak, M. B., Nagradova, N. K., and Muronetz, V. I. (1997) Arch. Biochem. Biophys., 345, 185–192.PubMedCrossRefGoogle Scholar
  17. 17.
    Grigorieva, J. A., Dainiak, M. B., Katrukha, A. G., and Muronetz, V. I. (1999) Arch. Biochem. Biophys., 369, 252–260.PubMedCrossRefGoogle Scholar
  18. 18.
    Arutyunova, E. I., Danshina, P. V., Domnina, L. V., Pleten, A. P., and Muronetz, V. I. (2003) Biochem. Biophys. Res. Commun., 307, 547–552.PubMedCrossRefGoogle Scholar
  19. 19.
    Domnina, L. V., Ivanova, O. Y., Cherniak, B. V., Skulachev, V. P., and Vasiliev, J. M. (2002) Biochemistry (Moscow), 67, 737–746.CrossRefGoogle Scholar
  20. 20.
    Rovensky, Y. A., Domnina, L. V., Ivanova, O. Y., and Vasiliev, J. M. (1999) J. Cell Sci., 112, 1273–1282.PubMedGoogle Scholar
  21. 21.
    Sidoti-de-Fraisse, C., Rincheval, V., Risler, Y., Mignotte, B., and Vayssiere, J. L. (1998) Oncogene, 17, 1639–1651.PubMedCrossRefGoogle Scholar
  22. 22.
    Olah, J., Tokesi, N., Vincze, O., Horvath, I., Lehotzky, A., Erdei, A., Szajli, E., Medzihradszky, K. F., Orosz, F., Kovacs, G. G., and Ovadi, J. (2006) FEBS Lett., 580, 5807–5814.PubMedCrossRefGoogle Scholar
  23. 23.
    Poglazov, B. F., and Livanova, N. B. (1986) Adv. Enzyme Regul., 25, 297–305.PubMedCrossRefGoogle Scholar
  24. 24.
    Minaschek, G., Groschel-Stewart, U., Blum, S., and Bereiter-Hahn, J. (1992) Eur. J. Cell Biol., 58, 418–428.PubMedGoogle Scholar
  25. 25.
    Waingeh, V. F., Gustafson, C. D., Kozliak, E. I., Lowe, S. L., Knull, H. R., and Thomasson, K. A. (2006) Biophys. J., 90, 1371–1384.PubMedCrossRefGoogle Scholar
  26. 26.
    Ouporov, I. V., Knull, H. R., Lowe, S. L., and Thomasson, K. A. (2001) J. Mol. Recognit., 14, 29–41.PubMedCrossRefGoogle Scholar
  27. 27.
    Tisdale, E. J., Kelly, C., and Artalejo, C. R. (2004) J. Biol. Chem., 279, 54046–54052.PubMedCrossRefGoogle Scholar
  28. 28.
    Muronetz, V. I., Wang, Z. X., Keith, T. J., Knull, H. R., and Srivastava, D. K. (1994) Arch. Biochem. Biophys., 313, 253–260.PubMedCrossRefGoogle Scholar
  29. 29.
    Pierce, A., Mirzaei, H., Muller, F., De Waal, E., Taylor, A. B., Leonard, S., Van Remmen, H., Regnier, F., Richardson, A., and Chaudhuri, A. (2008) J. Mol. Biol., 382, 1195–1210.PubMedCrossRefGoogle Scholar
  30. 30.
    Sen, N., Hara, M. R., Kornberg, M. D., Cascio, M. B., Bae, B. I., Shahani, N., Thomas, B., Dawson, T. M., Dawson, V. L., Snyder, S. H., and Sawa, A. (2008) Nat. Cell Biol., 10, 866–873.PubMedCrossRefGoogle Scholar
  31. 31.
    Chuang, D. M., Hough, C., and Senatorov, V. V. (2005) Annu. Rev. Pharmacol. Toxicol., 45, 269–290.PubMedCrossRefGoogle Scholar
  32. 32.
    Tarze, A., Deniaud, A., Le Bras, M., Maillier, E., Molle, D., Larochette, N., Zamzami, N., Jan, G., Kroemer, G., and Brenner, C. (2007) Oncogene, 26, 2606–2620.PubMedCrossRefGoogle Scholar
  33. 33.
    Colell, A., Ricci, J. E., Tait, S., Milasta, S., Maurer, U., Bouchier-Hayes, L., Fitzgerald, P., Guio-Carrion, A., Waterhouse, N. J., Li, C. W., Mari, B., Barbry, P., Newmeyer, D. D., Beere, H. M., and Green, D. R. (2007) Cell, 129, 983–997.PubMedCrossRefGoogle Scholar
  34. 34.
    Nakajima, H., Amano, W., Fujita, A., Fukuhara, A., Azuma, Y. T., Hata, F., Inui, T., and Takeuchi, T. (2007) J. Biol. Chem., 282, 26562–26574.PubMedCrossRefGoogle Scholar
  35. 35.
    Naletova, I., Schmalhausen, E., Kharitonov, A., Katrukha, A., Saso, L., Caprioli, A., and Muronetz, V. (2008) Biochim. Biophys. Acta, 1784, 2052–2058.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2013

Authors and Affiliations

  • E. I. Arutyunova
    • 1
  • L. V. Domnina
    • 1
  • A. A. Chudinova
    • 1
  • O. N. Makshakova
    • 1
  • D. Y. Arutyunov
    • 1
  • V. I. Muronetz
    • 1
    Email author
  1. 1.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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