Fludarabine nucleoside induces accumulations of p53, p63 and p73 in the nuclei of human B-lymphoid cell lines, with cytosolic and mitochondrial increases in p53
Article first published online: 29 JUN 2012
© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PROTEOMICS - Clinical Applications
Volume 6, Issue 5-6, pages 279–290, June 2012
How to Cite
Almazi, J. G., Mactier, S., Best, O. G., Crossett, B., Mulligan, S. P. and Christopherson, R. I. (2012), Fludarabine nucleoside induces accumulations of p53, p63 and p73 in the nuclei of human B-lymphoid cell lines, with cytosolic and mitochondrial increases in p53. Prot. Clin. Appl., 6: 279–290. doi: 10.1002/prca.201200003
- Issue published online: 29 JUN 2012
- Article first published online: 29 JUN 2012
- Accepted manuscript online: 29 MAY 2012 02:04AM EST
- Manuscript Accepted: 6 APR 2012
- Manuscript Revised: 16 MAR 2012
- Manuscript Received: 12 JAN 2012
- B-lymphoproliferative disorders;
Human Raji cells treated with fludarabine nucleoside (2-FaraA, 3 μM) undergo apoptosis with accumulation of p53 in the nuclei as multiple phosphorylated isoforms and C-terminal truncated derivatives. Changes induced by 2-FaraA in the levels of p53, p63 and p73 in the nuclear, cytosolic and mitochondrial fractions have been determined in four human B-lymphoid cell lines that are TP53-functional (Raji and IM9) and TP53-mutated (MEC1 and U266).
The B-lymphoid cell lines were treated with 2-FaraA (3 μM, 24 h, 48 h) and viability determined. Protein extracts of subcellular fractions from 2-FaraA-treated cells were analysed by 1D and 2D electrophoresis; multiple phosphorylated isoforms and truncated derivatives were identified by Western blots for p53, p63 and p73.
p53 and p63 were present in all three fractions, while p73 was only detected in nuclei. After treatment with 2-FaraA, nuclear p53, p63 and p73 accumulated as multiple phosphorylated isoforms and truncated derivatives. The association of p63 with mitochondria in human cells is novel.
Conclusions and clinical relevance
Comprehensive information on the subcellular distributions and responses of p53, p63 and p73 to 2-FaraA provides additional insight into mechanisms for induction of apoptosis in the treatment of B-lymphoproliferative disorders with fludarabine.