Allosteric effects mediate CHK2 phosphorylation of the p53 transactivation domain

Ashley Craig, Mary Scott, Lindsay Burch, Graeme Smith, Kathryn Ball, Ted Hupp

Research output: Contribution to journalArticlepeer-review

Abstract

The tumour suppressor p53 is a tetrameric protein that is phosphorylated in its BOX-I transactivation domain by checkpoint kinase 2 (CHK2) in response to DNA damage. CHK2 cannot phosphorylate small peptide fragments of p53 containing the BOX-I motif, indicating that undefined determinants in the p53 tetramer mediate CHK2 recognition. Two peptides derived from the DNA-binding domain of p53 bind to CHK2 and stimulate phosphorylation of full-length p53 at Thr 18 and Ser 20, thus identifying CHK2-docking sites. CHK2 can be fully activated in trans by the two p53 DNA-binding-domain peptides, and can phosphorylate BOX-I transactivation-domain fragments of p53 at Thr 18 and Ser 20. Although CHK2 has a basal Ser 20 kinase activity that is predominantly activated towards Thr 18, CHK1 has constitutive Thr 18 kinase activity that is predominantly activated in trans towards Ser 20. Cell division cycle 25C (CDC25C) phosphorylation by CHK2 is unaffected by the p53 DNA-binding-domain peptides. The CHK2-docking site in the BOX-V motif is the smallest of the two CHK2 binding sites, and mutating certain amino acids in the BOX-V peptide prevents CHK2 activation. A database search identified a p53 BOX-I-homology motif in p21(WAF1) and although CHK2 is inactive towards this protein, the p53 DNA-binding-domain peptides induce phosphorylation of p21(WAF1) at Ser 146. This provides evidence that CHK2 can be activated allosterically towards some substrates by a novel docking interaction, and identify a potential regulatory switch that may channel CHK2 into distinct signalling pathways in vivo.
Original languageEnglish
Pages (from-to)787-92
Number of pages6
JournalEMBO Reports
Volume4
Issue number8
DOIs
Publication statusPublished - Aug 2003

Keywords

  • Allosteric Regulation
  • Amino Acid Motifs
  • Binding Sites
  • Cell Cycle Proteins
  • Cell Line
  • Checkpoint Kinase 2
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins
  • Humans
  • Peptide Fragments
  • Phosphorylation
  • Protein Binding
  • Protein-Serine-Threonine Kinases
  • Signal Transduction
  • Transcriptional Activation
  • Tumor Suppressor Protein p53

Fingerprint

Dive into the research topics of 'Allosteric effects mediate CHK2 phosphorylation of the p53 transactivation domain'. Together they form a unique fingerprint.

Cite this