Cited by 972Cancer-related Molecular PathwaysDNA Repair MechanismsHeat shock proteins research

The human homologs of checkpoint kinases Chk1 and Cds1 (Chk2) phosphorylate p53 at multiple DNA damage-inducible sites.

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Shieh, Sheau-Yann, Ahn, Jinwoo, Tamai, Katsuyuki, Taya, Yoichi, Prives, Carol·2000·Genes & Development

Abstract

Upon DNA damage, the amino terminus of p53 is phosphorylated at a number of serine residues including S20, a site that is particularly important in regulating stability and function of the protein. Because no known kinase has been identified that can modify this site, HeLa nuclear extracts were fractionated and S20 phosphorylation was followed. We discovered that a S20 kinase activity copurifies with the human homolog of the Schizosaccharomyces pombe checkpoint kinase, Chk1 (hCHK1). We confirmed that recombinant hCHK1, but not a kinase-defective version of hCHK1, can phosphorylate p53 in vitro at S20. Additional inducible amino- and carboxy-terminal sites in p53 are also phosphorylated by hCHK1, indicating that this is an unusually versatile protein kinase. It is interesting that hCHK1 strongly prefers tetrameric to monomeric p53 in vitro, consistent with our observation that phosphorylation of amino-terminal sites in vivo requires that p53 be oligomeric. Regulation of the levels and activity of hCHK1 in transfected cells is directly correlated with the levels of p53; expression of either a kinase-defective hCHK1 or antisense hCHK1 leads to reduced levels of cotransfected p53, whereas overexpression of wild-type hCHK1 or the kinase domain of hCHK1 results in increased levels of expressed p53 protein. The human homolog of the second S. pombe checkpoint kinase, Cds1 (CHK2/hCds1), phosphorylates tetrameric p53 but not monomeric p53 in vitro at sites similar to those phosphorylated by hCHK1 kinase, suggesting that both checkpoint kinases can play roles in regulating p53 after DNA damage.

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Crossref · 10.1101/gad.14.3.289

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  • hChk2 phosphorylates p53 at Ser20.

    “and hChk2, in turn, phosphorylates yet another amino-terminal Ser residue (Ser 20) in p53”

    From Cell cycle checkpoint signaling through the ATM and ATR kinases (2001) · cites this paper as Shieh et al. 2000

  • These phosphorylation events disrupt p53-Mdm2 and Mdm2-DAXX interactions and promote degradation of Mdm2 and Mdm4.

    “These phosphorylation events contribute to disrupting the p53-Mdm2 (Chehab et al., 2000; Shieh et al., 2000), and Mdm2-DAXX (Tang et al., 2013) interactions, as well as to the degradation of Mdm2 and Mdm4 (Stommel and Wahl, 2004; Chen et al., 2005; Pereg et al., 2005, 2006; LeBron et al., 2006).”

    From USP7 Is a Master Regulator of Genome Stability (2020) · cites this paper as Shieh et al., 2000

  • p53 is phosphorylated at Ser20 by Chk2 after IR, resulting in p53's dissociation from Mdm2 and its subsequent stabilization.

    “p53 is phosphorylated at Ser20 by the checkpoint kinase Chk2 after IR, which results in p53's dissociation from Mdm2 and its subsequent stabilization”

    From USP10 Regulates p53 Localization and Stability by Deubiquitinating p53 (2010) · cites this paper as Shieh et al., 2000

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Research works printed but not linked: 1

  1. Chehab, N.H., A. Malikzay, E.S. Stavridi, and T.D. Halazonetis. 1999. Phosphorylation of Ser20 mediates stabilization of hu- man p53 in response to DNA damage. Proc. Natl. Acad. Sci. 96: 13777-13782.

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Authors
Shieh, S.-Y., Ahn, J., Tamai, K., Taya, Y., & Prives, C.
Affiliations
Seisen University (JP)
Venue
Genes & Development
Published
2000-02-01
Volume
14
Issue
3
Pages
289-300
ISSN
  • 0890-9369 (ISSN-L)
  • 1549-5477
OpenAlex
W2120921507
Cited by
972
References
84
Citations by year
2026: 11 · 2025: 4 · 2024: 15 · 2023: 22 · 2022: 21 · 2021: 21 · 2020: 28 · 2019: 26 · 2018: 22 · 2017: 19 · 2016: 22 · 2015: 24 · 2014: 39 · 2013: 38 · 2012: 42
Type
posted content
Language
en
Open access
Locations
  • Genes & Development (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Princess Takamatsu Cancer Research Fund
  • National Institutes of Health (CA58316)
  • National Cancer Institute (R37CA058316, R01CA058316, CA 58316)
Topics
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
Keywords
p53 phosphorylation, Chk1, Chk2, checkpoint kinase, DNA damage response, p53 oligomerization, p53 stabilization, Schizosaccharomyces pombe, kinase activity
MeSH terms
Checkpoint Kinase 1, DNA Damage, HeLa Cells, Humans, Phosphorylation, Protein Kinases, Recombinant Proteins, Blotting, Western, Tumor Suppressor Protein p53, Protein Serine-Threonine Kinases, Protein Structure, Tertiary, Schizosaccharomyces pombe Proteins, Checkpoint Kinase 2

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