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Carcinogenesis, Vol. 21, No. 2, 153-160, February 2000
© 2000 Oxford University Press


Cancer Biology

Expression of the p53 homologue p63{alpha} and {Delta}Np63{alpha} in normal and neoplasticcells

Peter A. Hall1, Sandra J. Campbell, Mary O'neill, Daniel J. Royston, Karin Nylander, Frank A. Carey and Neil M. Kernohan

Department of Molecular and Cellular Pathology, University of Dundee, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK

A burgeoning family of p53-related genes have been described recently, including p73 and p63. Both these genes encode proteins with many similarities to p53 but also with the potential for forming a range of related species by alternative promoter usage and alternative splicing. In order to begin the characterization of p63, we generated a polyclonal serum (designated SC1) that recognizes the C-terminus of p63{alpha}. We have shown that this reagent recognizes p63{alpha} but not p53 nor p73. By western blot analysis both p63{alpha} and the N-terminal truncated form of p63{alpha} ({Delta}Np63{alpha}) were found in a range of cell lines. Similar immunoblot analysis of tissues reveals considerable complexity with at least four SC1-immunoreactive isoforms being identified. In immunohistological studies SC1 immunoreactivity is widely detectable, being predominantly associated with proliferative compartments in epithelia. However, non-proliferative populations can also show SC1 immunostaining. No simple relationship between the isoforms identified by immunoblotting of tissue lysates and the tissue immunostaining characteristics was identified. A previously unrecognized species intermediate in mobility between p63{alpha} and {Delta}Np63{alpha} was found in several tissues, including nerve and peripheral blood lymphocytes. Interestingly, there is suppression of p63{alpha} expression in HaCat cells in a time- and concentration-dependent manner after UV and MMS treatment. Our data provide further information about the complexity of p63 and the SC1 serum will prove to be a useful tool in further studies of this p53 homologue.

Abbreviations: LMS, limb mammary syndrome; PBST, phosphate-buffered saline and 0.1% Tween 20.


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