DNA damage triggers DRB-resistant phosphorylation of human p53 at the CK2 site.
The sequence-specific DNA binding activity of p53 is negatively regulated by a C-terminal domain whose phosphorylation in vitro can activate the latent DNA binding function of the protein. The DNA binding activity of p53 is a core component of its stress-activated transcription function, yet it is not yet clear whether phosphorylation within the C-terminal domain plays a role in the p53 damage response in vivo. As the casein kinase 2 (CK2) site at serine 392 is the C-terminal phosphorylation motif that exhibits the most pronounced conservation at the primary amino acid level, we have focused on determining whether the CK2 site is modified in vivo and whether radiation effects the extent of that phosphorylation. Using antibodies that can detect serine 392-phosphorylation of p53, we demonstrate that UV radiation can trigger extensive phosphorylation at the CK2 site. The CK2 inhibitor, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), can partially inhibit the UV-induced phosphorylation at serine 392, suggesting that CK2 is one of the major serine 392-kinases. However, a striking increase in UV-induced serine 392 phosphorylation and p53 transactivation function at higher levels of DRB suggests that a DRB-resistant/stress-activated pathway may target serine 392 in vivo. These data demonstrate that radiation-induced phosphorylation of p53 can occur in vivo at serine 392 and implicate a CK2-independent signal cascade that can function to modulate serine 392 phosphorylation in cells.
Blaydes
J P
JP
Department of Molecular and Cellular Pathology, Ninewells Medical School, The University of Dundee, Scotland, UK.
Hupp
T R
TR
D001665:Binding Sites / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D047390:Casein Kinase II; D004249:DNA Damage / Q000187:drug effects* / Q000528:radiation effects; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm / Q000235:genetics / Q000528:radiation effects; D006801:Humans; D010766:Phosphorylation / Q000187:drug effects / Q000528:radiation effects; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012694:Serine / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000528:radiation effects; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000378:metabolism* / Q000528:radiation effects; D014466:Ultraviolet Rays
D016159:Tumor Suppressor Protein p53; D012694:Serine; D004004:Dichlororibofuranosylbenzimidazole; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases