JBPC  Vol.2 No.4 , November 2011
Different behavior of protein B23/nucleophosmin and UBF in HeLa cells during apoptosis
ABSTRACT
The behavior of UBF (upstream binding factor) and nucleophosmin in HeLa and HeLa-Bcl-2 cells during apoptosis induced by TNF-α, emetine, and their mixture was investigated. A pronounced apoptosis was achieved only in HeLa cells treated with a mixture of the inducers. Immunoblotting analysis of UBF and nucleophosmin in samples containing different portions of cells with apoptotic nuclei was carried out. It showed that UBF was proteolytically cleaved giving a stable 76-kDa fragment. Increasing content of the fragment during apoptosis correlated with the level of cells containing apoptotic nuclei and with a decrease in the content of full-sized UBF. Determination of N- and C-terminal sequences of UBF and 76-kDa fragment allowed us not only to characterize UBF at the protein level, but also to describe the site of the apoptosis-specific proteolysis. Nucleophosmin did not undergo proteolytic cleavage during apoptosis and its content was unchanged even in a sample containing 100% of cells with apoptotic nuclei. However in cells reached terminal stages of apoptosis, the balance between mono- and oligomeric forms of nucleophosmin changed due to depletion of monomeric forms and appearance of two additional oligomeric forms with lower molecular weight.

Cite this paper
nullVladimirova, N. and Potapenko, N. (2011) Different behavior of protein B23/nucleophosmin and UBF in HeLa cells during apoptosis. Journal of Biophysical Chemistry, 2, 422-429. doi: 10.4236/jbpc.2011.24049.
References
[1]   Horky, M., Wurzer, G., Kotala, V., Anton, M., Vojtesek, B., Vacha, J. and Wesierska-Gadek, J. (2001) Segregation of nucleolar components coincides with caspase-3 activa- tion in cisplatin-treated HeLa cells. Journal of Cell Sci- ence, 114, 663-670.

[2]   Liu, W.H. and Yung, B.Y. (1998) Mortalization of human promyelocytic leukemia HL-60 cells to be more suscepti- ble to sodium butyrate-induced apoptosis and inhibition of telomerase activity by down-regulation of nucleo- phosmin/B23. Oncogene, 17, 3055-3064. doi:10.1038/sj.onc.1202234

[3]   Ulanet, D.B., Torbenson, M., Dang, C.V., Casciola-Rosen, L. and Rosen, A. (2003) Unique conformation of cancer autoantigen B23 in hepatoma: A mechanism for specifici- ty in the autoimmune response. Proceedings of the Na- tional Academy of Sciences of the United States of Ame- rica, 100, 12361-12366. doi:10.1073/pnas.2035245100

[4]   Casiano, C.A. and Tan, E.M. (1996) Antinuclear autoanti- bodies: Probes for defining proteolytic events associated with apoptosis. Molecular Biology Reports, 23, 211-216.

[5]   Martelli, A.M., Robuffo, I., Bortul, R., Ochs, R.L., Luchetti, F., Cocco, L., Zweyer, M., Bareggi, R. and Falcieri, E. (2000) Behavior of nucleolar proteins during the course of apoptosis in camptothecin-treated HL60 cells. Journal of Cellular Biochemistry, 78, 264-277. doi:10.1002/(SICI)1097-4644(20000801)78:2<264::AID-JCB9>3.3.CO;2-V

[6]   Grisendi, S., Mecucci, C., Falini, B. and Pandolfi, P.P. (2006) Nucleophosmin and cancer. Nature Reviews. Can- cer, 6, 493-505. doi:10.1038/nrc1885

[7]   Patterson, S.D., Grossman, J.S., D’Andrea, P. and Latter, G.I. (1995) Reduced numatrin/B23/nucleophosmin labe- ling in apoptotic Jurkat T-lymphoblasts. The Journal of Biological Chemistry, 270, 9429-9436.

[8]   Torres-Montaner, A., Bolivar, J., Astola, A., Gimenez- Mas, J.A., Brieva, J.A. and Valdivia, M.M. (2000) Im- munohistochemical detection of ribosomal transcription factor UBF and AgNOR staining identify apoptotic events in neoplastic cells of Hodgkin’s disease and in other lymphoid cells. The Journal of Histochemistry and Cy- tochemistry: Official Journal of the Histochemistry Soci- ety, 48, 1521-1530. doi:10.1177/002215540004801109

[9]   Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry, 193, 265- 275.

[10]   Vladimirova, N.M., Potapenko, N.A., Sachs, G. and Mo- dyanov, N.N. (1995) Determination of the sidedness of the carboxy-terminus of the Na+/K(+)ATPase alpha- subunit using lactoperoxidase iodination. Biochimica et Biophysica Acta, 1233, 175-184.

[11]   Sautkina, E.N., Potapenko, N.A. and Vladimirova, N.M. (2006) State of nucleolar proteins B23/nucleophosmin and UBF in HeLa cells during apoptosis induced by tu- mor necrosis factor. Biochemistry (Moscow), 71, 634- 643. doi:10.1134/S0006297906060071

[12]   Vladimirova, N.M., Sautkina, E.N., Tabdanov, E.D., et al. (2004) Identification and characterization of monomeric and oligomeric isoforms of a major nucleolar protein B23/nucleophosmin in HeLa cells. Biological Membrany (Moscow), 21, 94-101.

[13]   Rath, P.C. and Aggarwal, B.B. (1999) TNF-induced sig- naling in apoptosis. Journal of Clinical Immunology, 19, 350-364.

[14]   Schweighoffer, T., Schweighoffer, E., Sviresheva, M., Apati, A., Balogh, A., Csuka, I., Luat, N.N. and Banfalvi, G. (1993) Oncogene expression in thymocytes after em- etine treatment of mice. Cellular and Molecular Biology Research, 39, 647-656.

[15]   Lazebnik, Y.A., Cole, S., Cooke, C.A., Nelson, W.G. and Earnshaw, W.C. (1993) Nuclear events of apoptosis in vitro in cell-free mitotic extracts: A model system for analysis of the active phase of apoptosis. The Journal of Cell Biology, 123, 7-22. doi:10.1083/jcb.123.1.7

[16]   Domnina, L.V., Ivanova, O.Y., Cherniak, B.V., Skulachev, V. P. and Vasiliev, J.M. (2002) Effects of the inhibitors of dynamics of cytoskeletal structures on the development of apoptosis induced by the tumor necrosis factor. Biochemistry (Moscow), 67, 737-746.

[17]   Robaye, B., Doskeland, A.P., Suarez-Huerta, N., Doske- land, S.O. and Dumont, J.E. (1994) Apoptotic cell death analyzed at the molecular level by two-dimensional gel electrophoresis. Electrophoresis, 15, 503-510. doi:10.1002/elps.1150150168

[18]   Jantzen, H.M., Admon, A., Bell, S.P. and Tjian, R. (1990) Nucleolar transcription factor hUBF contains a DNA- binding motif with homology to HMG proteins. Nature, 344, 830-836. doi:10.1038/344830a0

[19]   Chan, P.K. and Chan, F.Y. (1995) Nucleophosmin/B23 (NPM) oligomer is a major and stable entity in HeLa cells. Biochimica et Biophysica acta, 1262, 37-42. doi:10.1016/0167-4781(95)00044-H

[20]   Vladimirova, N.M., Lobanova, N.V. and Potapenko, N.A. (2010) State of oncomarker protein B23/nucleophosmin in HeLa cells. Biochemistry (Moscow), 75, 851-860. doi:10.1134/S0006297910070060

 
 
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