Molecular basis of S100 proteins interacting with the p53 homologs p63 and p73.
S100 proteins modulate p53 activity by interacting with its tetramerization (p53TET, residues 325-355) and transactivation (residues 1-57) domains. In this study, we characterized biophysically the binding of S100A1, S100A2, S100A4, S100A6 and S100B to homologous domains of p63 and p73 in vitro by fluorescence anisotropy, analytical ultracentrifugation and analytical gel filtration. We found that S100A1, S100A2, S100A4, S100A6 and S100B proteins bound different p63 and p73 tetramerization domain variants and naturally occurring isoforms with varying affinities in a calcium-dependent manner. Additional interactions were observed with peptides derived from the p63 and p73 N-terminal transactivation domains. Importantly, S100 proteins bound p63 and p73 with different affinities in their different oligomeric states, similarly to the differential modes of binding to p53. On the basis of our data, we hypothesize that S100 proteins regulate the oligomerization state of all three p53 family members and their isoforms, with a potential physiological relevance in developmental and disease-related processes. The regulation of the p53 family by S100 is complicated and depends on the target preference of each individual S100 protein, the concentration of the proteins and calcium, as well as the splicing variation of p63 or p73. Our results outlining the complexity of the interaction should be considered when studying the functional effects of S100 proteins in their biological context.
MRC Centre for Protein Engineering, Hills Road, Cambridge, UK.
J
J
van Dieck
T
T
Brandt
D P
DP
Teufel
D B
DB
Veprintsev
A C
AC
Joerger
A R
AR
Fersht
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002850:Chromatography, Gel; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D005454:Fluorescence Polarization; D006801:Humans; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D009418:S100 Proteins / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D015533:Transcriptional Activation; D000072160:Tumor Protein p73; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry*; D025521:Tumor Suppressor Proteins / Q000737:chemistry* / Q000378:metabolism*; D014461:Ultracentrifugation
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D009418:S100 Proteins; C000606460:TP73 protein, human; D000072160:Tumor Protein p73; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D002118:Calcium