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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

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• TITLE: Different behavior of protein B23/nucleophosmin and UBF in HeLa cells during apoptosis

  AUTHORS: Natalia M. Vladimirova, Natalia A. Potapenko

  KEYWORDS: TNF-α Induced Apoptosis; Proteolysis of UBF; Monomer-Oligomer State of Nucleophosmin

  JOURNAL NAME: Journal of Biophysical Chemistry, Vol.2 No.4, November 9, 2011

  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.