title: Concerted regulation of wild-type p53 nuclear accumulation and activation by S100B and calcium-dependent protein kinase C.
abstract: The calcium ionophore ionomycin cooperates with the S100B protein to rescue a p53-dependent G(1) checkpoint control in S100B-expressing mouse embryo fibroblasts and rat embryo fibroblasts (REF cells) which express the temperature-sensitive p53Val135 mutant (C. Scotto, J. C. Deloulme, D. Rousseau, E. Chambaz, and J. Baudier, Mol. Cell. Biol. 18:4272-4281, 1998). We investigated in this study the contributions of S100B and calcium-dependent PKC (cPKC) signalling pathways to the activation of wild-type p53. We first confirmed that S100B expression in mouse embryo fibroblasts enhanced specific nuclear accumulation of wild-type p53. We next demonstrated that wild-type p53 nuclear translocation and accumulation is dependent on cPKC activity. Mutation of the five putative cPKC phosphorylation sites on murine p53 into alanine or aspartic residues had no significant effect on p53 nuclear localization, suggesting that the cPKC effect on p53 nuclear translocation is indirect. A concerted regulation by S100B and cPKC of wild-type p53 nuclear translocation and activation was confirmed with REF cells expressing S100B (S100B-REF cells) overexpressing the temperature-sensitive p53Val135 mutant. Stimulation of S100B-REF cells with the PKC activator phorbol ester phorbol myristate acetate (PMA) promoted specific nuclear translocation of the wild-type p53Val135 species in cells positioned in early G(1) phase of the cell cycle. PMA also substituted for ionomycin in the mediating of p53-dependent G(1) arrest at the nonpermissive temperature (37.5 degrees C). PMA-dependent growth arrest was linked to the cell apoptosis response to UV irradiation. In contrast, growth arrest mediated by a temperature shift to 32 degrees C protected S100B-REF cells from apoptosis. Our results suggest a model in which calcium signalling, linked with cPKC activation, cooperates with S100B to promote wild-type p53 nuclear translocation in early G(1) phase and activation of a p53-dependent G(1) checkpoint control.
authors.affiliation: Département de Biologie Moléculaire et Structurale du CEA, DBMS-BRCE INSERM Unité 244, 38054 Grenoble Cedex 9, France.
authors.forename: C
authors.initials: C
authors.lastname: Scotto
authors.forename: C
authors.initials: C
authors.lastname: Delphin
authors.forename: J C
authors.initials: JC
authors.lastname: Deloulme
authors.forename: J
authors.initials: J
authors.lastname: Baudier
mesh_terms: D000818:Animals; D001692:Biological Transport; D020013:Calcium Signaling; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002227:Carbazoles / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism*; D016193:G1 Phase; D007211:Indoles / Q000494:pharmacology; D051379:Mice; D009414:Nerve Growth Factors / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D013755:Tetradecanoylphorbol Acetate; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D002135:Calcium-Binding Proteins; D002227:Carbazoles; D007211:Indoles; D009414:Nerve Growth Factors; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; C579159:S100b protein, mouse; C579158:S100b protein, rat; D016159:Tumor Suppressor Protein p53; C081021:Go 6976; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate