title: Mechanistic differences in the transcriptional activation of p53 by 14-3-3 isoforms.
abstract: p53 maintains genome integrity by initiating the transcription of genes involved in cell-cycle arrest, senescence, apoptosis and DNA repair. The activity of p53 is regulated by both post-translational modifications and protein-protein interactions. p53 that has been phosphorylated at S366, S378 and T387 binds 14-3-3 proteins in vitro. Here, we show that these sites are potential 14-3-3 binding sites in vivo. Epsilon (epsilon) and gamma (gamma) isoforms required phosphorylation at either of these sites for efficient interaction with p53, while for sigma (sigma) and tau (tau) these sites are dispensable. Further, sigma and tau bound more weakly to p53 C-terminal phosphopeptides than did epsilon and gamma. However, the four isoforms bound tightly to di-phosphorylated p53 C-terminal peptides than did the mono-phosphorylated counterparts. Interestingly, all the isoforms studied transcriptionally activated wild-type p53. sigma and tau stabilized p53 levels in cells, while epsilon and gamma stimulated p53-DNA binding activity in vitro. Overall, the results suggest that structurally and functionally similar 14-3-3 isoforms may exert their regulatory potential on p53 through different mechanisms. We discuss the isoform-specific roles of 14-3-3 in p53 stabilization and activation of specific-DNA binding.
authors.affiliation: MRC Centre for Protein Engineering, Hills Road, Cambridge, CB20QH, UK.
authors.forename: Sridharan
authors.initials: S
authors.lastname: Rajagopalan
authors.forename: Robert S
authors.initials: RS
authors.lastname: Sade
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Townsley
authors.forename: Alan R
authors.initials: AR
authors.lastname: Fersht
mesh_terms: D048948:14-3-3 Proteins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D006801:Humans; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013816:Thermodynamics; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000378:metabolism*
chemical_list: D048948:14-3-3 Proteins; D020033:Protein Isoforms; D016159:Tumor Suppressor Protein p53; D004247:DNA