Expression of the p53 homologue p63alpha and deltaNp63alpha in normal and neoplastic cells.
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 p63alpha. We have shown that this reagent recognizes p63alpha but not p53 nor p73. By western blot analysis both p63alpha and the N-terminal truncated form of p63alpha (DeltaNp63alpha) 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 p63alpha and DeltaNp63alpha was found in several tissues, including nerve and peripheral blood lymphocytes. Interestingly, there is suppression of p63alpha 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.
Hall
P A
PA
Department of Molecular and Cellular Pathology, University of Dundee, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK.
Campbell
S J
SJ
O'neill
M
M
Royston
D J
DJ
Nylander
K
K
Carey
F A
FA
Kernohan
N M
NM
D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000032:analysis; D015153:Blotting, Western; D002455:Cell Division; D004268:DNA-Binding Proteins; D016147:Genes, Tumor Suppressor; D006801:Humans; D007106:Immune Sera; D008565:Membrane Proteins; D016685:Mitomycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000032:analysis; D009928:Organ Specificity; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000235:genetics; D020033:Protein Isoforms / Q000096:biosynthesis* / Q000235:genetics; D015534:Trans-Activators; D014157:Transcription Factors; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000528:radiation effects; D025521:Tumor Suppressor Proteins; D014466:Ultraviolet Rays
D001798:Blood Proteins; C081876:CKAP4 protein, human; D004268:DNA-Binding Proteins; D007106:Immune Sera; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D020033:Protein Isoforms; C115051:TP63 protein, human; D015534:Trans-Activators; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D016685:Mitomycin