Phosphorylation at carboxyl-terminal S373 and S375 residues and 14-3-3 binding are not required for mouse p53 function.
gamma-Irradiation-mediated ataxia telangiectasia mutated (ATM)-dependent dephosphorylation of serine 376 (S376) at the carboxyl terminus of human p53 results in the exposure of the 14-3-3 consensus-binding site, which includes serine 378 (S378). 14-3-3 binding potentiates p53's DNA-binding ability and causes G(1) arrest. Moreover,endoplasmic reticulum stress-mediated S376 phosphorylation was shown to localize human p53 in the cytoplasm. Although many functions are conserved between human and mouse p53, the functional relevance of S376 and S378 mouse equivalents is not clear. We report here that gamma-irradiation does not lead to 14-3-3 binding to mouse p53. Mouse p53 mutants, such as S373A/D (the equivalent of human S376), S375A/D (the equivalent of human S378), and combinatorial double mutants, were not impaired in their ability to transactivate p53-dependent target genes and were capable of inducing G(1) arrest as efficiently as wild-type p53. Consistently, all mutant p53s were as potent as wild-type mouse p53 in inhibiting cellular colony formation. Furthermore, mouse S373A/D mutants were not defective in cytoplasmic localization in response to endoplasmic reticulum stress. Together, the data suggest that despite a high homology with human p53, neither phosphorylation status at S373 and S375 nor 14-3-3 binding may be a critical event for mouse p53 to be functional.
Lee
Ming Kei
MK
Laboratory of Molecular Carcinogenesis, National Cancer Center, Singapore 169610, Singapore.
Sabapathy
Kanaga
K
D048948:14-3-3 Proteins / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D001665:Binding Sites / Q000235:genetics / Q000502:physiology; D045744:Cell Line, Tumor / Q000378:metabolism / Q000528:radiation effects; D002478:Cells, Cultured / Q000378:metabolism / Q000528:radiation effects; D005347:Fibroblasts / Q000378:metabolism / Q000528:radiation effects; D016193:G1 Phase; D005720:Gamma Rays; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D010768:Phosphoserine / Q000737:chemistry*; D011485:Protein Binding / Q000528:radiation effects; D011499:Protein Processing, Post-Translational; D016415:Sequence Alignment; D017385:Sequence Homology; D013045:Species Specificity; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000172:deficiency / Q000502:physiology*
D048948:14-3-3 Proteins; D016159:Tumor Suppressor Protein p53; D010768:Phosphoserine