Home LiteratureArticle Details
PMID: 10673500 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Chk2/hCds1 functions as a DNA damage checkpoint in G(1) by stabilizing p53.

Genes & development ·Vol. 14 ·No. 3 ·2000-02-01 ·Pages 278-88

Chehab NH, Malikzay A, Appel M, Halazonetis TD

Abstract

Chk2/hcds1, the human homolog of the Saccharomyces cerevisiae RAD53/SPK1 and Schizosaccharomyces pombe cds1 DNA damage checkpoint genes, encodes a protein kinase that is post-translationally modified after DNA damage. Like its yeast homologs, the Chk2/hCds1 protein phosphorylates Cdc25C in vitro, suggesting that it arrests cells in G(2) in response to DNA damage. We expressed Chk2/hCds1 in human cells and analyzed their cell cycle profile. Wild-type, but not catalytically inactive, Chk2/hCds1 led to G(1) arrest after DNA damage. The arrest was inhibited by cotransfection of a dominant-negative p53 mutant, indicating that Chk2/hCds1 acted upstream of p53. In vitro, Chk2/hCds1 phosphorylated p53 on Ser-20 and dissociated preformed complexes of p53 with Mdm2, a protein that targets p53 for degradation. In vivo, ectopic expression of wild-type Chk2/hCds1 led to increased p53 stabilization after DNA damage, whereas expression of a dominant-negative Chk2/hCds1 mutant abrogated both phosphorylation of p53 on Ser-20 and p53 stabilization. Thus, in response to DNA damage, Chk2/hCds1 stabilizes the p53 tumor suppressor protein leading to cell cycle arrest in G(1).

MeSH Terms
Checkpoint Kinase 2 DNA Damage G1 Phase Humans Nuclear Proteins Phosphorylation Protein Kinases Protein Serine-Threonine Kinases/metabolism Proto-Oncogene Proteins/metabolism Proto-Oncogene Proteins c-mdm2 Schizosaccharomyces pombe Proteins Tumor Cells, Cultured Tumor Suppressor Protein p53/metabolism
Chemicals
Nuclear Proteins Proto-Oncogene Proteins Schizosaccharomyces pombe Proteins Tumor Suppressor Protein p53 MDM2 protein, human Proto-Oncogene Proteins c-mdm2 Protein Kinases Checkpoint Kinase 2 CHEK2 protein, human Cds1 protein, S pombe Protein Serine-Threonine Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Chehab N H
Department of Molecular Genetics, The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
Malikzay A
Appel M
Halazonetis T D
References (77)
77 references, click to expand
  1. Separate pathways for p53 induction by ionizing radiation and N-(phosphonoacetyl)-L-aspartate.
    Cancer Res. 1996 Aug 15;56(16):3659-62 PMID: 8706003
  2. Structure-based rescue of common tumor-derived p53 mutants.
    Nat Med. 1996 Oct;2(10):1143-6 PMID: 8837616
  3. RAD9 and DNA polymerase epsilon form parallel sensory branches for transducing the DNA damage checkpoint signal in Saccharomyces cerevisiae.
    Genes Dev. 1996 Oct 15;10(20):2632-43 PMID: 8895664
  4. Structure of the MDM2 oncoprotein bound to the p53 tumor suppressor transactivation domain.
    Science. 1996 Nov 8;274(5289):948-53 PMID: 8875929
  5. Cell cycle checkpoints: preventing an identity crisis.
    Science. 1996 Dec 6;274(5293):1664-72 PMID: 8939848
  6. The Schizosaccharomyces pombe rad3 checkpoint gene.
    EMBO J. 1996 Dec 2;15(23):6641-51 PMID: 8978690
  7. p53, the cellular gatekeeper for growth and division.
    Cell. 1997 Feb 7;88(3):323-31 PMID: 9039259
  8. The structural basis for spectral variations in green fluorescent protein.
    Nat Struct Biol. 1997 May;4(5):361-5 PMID: 9145105
  9. Mdm2 promotes the rapid degradation of p53.
    Nature. 1997 May 15;387(6630):296-9 PMID: 9153395
  10. Regulation of p53 stability by Mdm2.
    Nature. 1997 May 15;387(6630):299-303 PMID: 9153396
  11. p53 transactivation and protein accumulation are independently regulated by UV light in different phases of the cell cycle.
    Mol Cell Biol. 1997 Jun;17(6):3074-80 PMID: 9154806
  12. The p53 activation and apoptosis induced by DNA damage are reversibly inhibited by salicylate.
    Oncogene. 1997 May 29;14(21):2503-10 PMID: 9191050
  13. Cdc25 mitotic inducer targeted by chk1 DNA damage checkpoint kinase.
    Science. 1997 Sep 5;277(5331):1495-7 PMID: 9278510
  14. Linkage of ATM to cell cycle regulation by the Chk2 protein kinase.
    Science. 1998 Dec 4;282(5395):1893-7 PMID: 9836640
  15. ATM associates with and phosphorylates p53: mapping the region of interaction.
    Nat Genet. 1998 Dec;20(4):398-400 PMID: 9843217
  16. A p53-dependent S-phase checkpoint helps to protect cells from DNA damage in response to starvation for pyrimidine nucleotides.
    Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14775-80 PMID: 9843965
  17. p53 sites acetylated in vitro by PCAF and p300 are acetylated in vivo in response to DNA damage.
    Mol Cell Biol. 1999 Feb;19(2):1202-9 PMID: 9891054
  18. A human homologue of the checkpoint kinase Cds1 directly inhibits Cdc25 phosphatase.
    Curr Biol. 1999 Jan 14;9(1):1-10 PMID: 9889122
  19. A role for ATR in the DNA damage-induced phosphorylation of p53.
    Genes Dev. 1999 Jan 15;13(2):152-7 PMID: 9925639
  20. An inhibitor of nuclear export activates the p53 response and induces the localization of HDM2 and p53 to U1A-positive nuclear bodies associated with the PODs.
    Exp Cell Res. 1999 May 1;248(2):457-72 PMID: 10222137
  21. Basis for the checkpoint signal specificity that regulates Chk1 and Cds1 protein kinases.
    Mol Cell Biol. 1999 Jun;19(6):4262-9 PMID: 10330167
  22. Change in oligomerization specificity of the p53 tetramerization domain by hydrophobic amino acid substitutions.
    Protein Sci. 1999 Sep;8(9):1773-9 PMID: 10493578
  23. Cdc7p-Dbf4p kinase binds to chromatin during S phase and is regulated by both the APC and the RAD53 checkpoint pathway.
    EMBO J. 1999 Oct 1;18(19):5334-46 PMID: 10508166
  24. Many faces of ATM: eighth international workshop on ataxia-telangiectasia.
    Biochim Biophys Acta. 1999 Oct 29;1424(2-3):R45-55 PMID: 10528155
  25. Role of human Cds1 (Chk2) kinase in DNA damage checkpoint and its regulation by p53.
    J Biol Chem. 1999 Oct 29;274(44):31463-7 PMID: 10531348
  26. Control of the DNA damage checkpoint by chk1 and rad53 protein kinases through distinct mechanisms.
    Science. 1999 Nov 5;286(5442):1166-71 PMID: 10550056
  27. Regulation of p53 function and stability by phosphorylation.
    Mol Cell Biol. 1999 Mar;19(3):1751-8 PMID: 10022862
  28. Loss of normal G1 checkpoint control is an early step in carcinogenesis, independent of p53 status.
    Cancer Res. 1999 Mar 1;59(5):1008-14 PMID: 10070956
  29. A human Cds1-related kinase that functions downstream of ATM protein in the cellular response to DNA damage.
    Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3745-50 PMID: 10097108
  30. Critical role for Ser20 of human p53 in the negative regulation of p53 by Mdm2.
    EMBO J. 1999 Apr 1;18(7):1805-14 PMID: 10202144
  31. DNA damage-inducible phosphorylation of p53 at N-terminal sites including a novel site, Ser20, requires tetramerization.
    EMBO J. 1999 Apr 1;18(7):1815-23 PMID: 10202145
  32. DNA damage induced p53 stabilization: no indication for an involvement of p53 phosphorylation.
    Oncogene. 1999 Mar 4;18(9):1723-32 PMID: 10208433
  33. Phosphorylation of Ser-20 mediates stabilization of human p53 in response to DNA damage.
    Proc Natl Acad Sci U S A. 1999 Nov 23;96(24):13777-82 PMID: 10570149
  34. Heterozygous germ line hCHK2 mutations in Li-Fraumeni syndrome.
    Science. 1999 Dec 24;286(5449):2528-31 PMID: 10617473
  35. The human homologs of checkpoint kinases Chk1 and Cds1 (Chk2) phosphorylate p53 at multiple DNA damage-inducible sites.
    Genes Dev. 2000 Feb 1;14(3):289-300 PMID: 10673501
  36. UV irradiation stimulates levels of p53 cellular tumor antigen in nontransformed mouse cells.
    Mol Cell Biol. 1984 Sep;4(9):1689-94 PMID: 6092932
  37. Primary structure and functional expression from complementary DNA of a human interleukin-1 receptor antagonist.
    Nature. 1990 Jan 25;343(6256):341-6 PMID: 2137201
  38. Participation of p53 protein in the cellular response to DNA damage.
    Cancer Res. 1991 Dec 1;51(23 Pt 1):6304-11 PMID: 1933891
  39. The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation.
    Cell. 1992 Jun 26;69(7):1237-45 PMID: 1535557
  40. Wild-type p53 is a cell cycle checkpoint determinant following irradiation.
    Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7491-5 PMID: 1323840
  41. A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia.
    Cell. 1992 Nov 13;71(4):587-97 PMID: 1423616
  42. Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA-damaging agents.
    Oncogene. 1993 Feb;8(2):307-18 PMID: 8426740
  43. p53 is required for radiation-induced apoptosis in mouse thymocytes.
    Nature. 1993 Apr 29;362(6423):847-9 PMID: 8479522
  44. Thymocyte apoptosis induced by p53-dependent and independent pathways.
    Nature. 1993 Apr 29;362(6423):849-52 PMID: 8479523
  45. Differential induction of transcriptionally active p53 following UV or ionizing radiation: defects in chromosome instability syndromes?
    Cell. 1993 Nov 19;75(4):765-78 PMID: 8242748
  46. Ionizing radiation and UV induction of p53 protein by different pathways in ataxia-telangiectasia cells.
    Oncogene. 1993 Dec;8(12):3307-12 PMID: 8247533
  47. The p53-dependent G1 cell cycle checkpoint pathway and ataxia-telangiectasia.
    Cancer Res. 1994 Oct 1;54(19):5054-8 PMID: 7923116
  48. A single ataxia telangiectasia gene with a product similar to PI-3 kinase.
    Science. 1995 Jun 23;268(5218):1749-53 PMID: 7792600
  49. Conservation of the Chk1 checkpoint pathway in mammals: linkage of DNA damage to Cdk regulation through Cdc25.
    Science. 1997 Sep 5;277(5331):1497-501 PMID: 9278511
  50. p53 protein stability in tumour cells is not determined by mutation but is dependent on Mdm2 binding.
    Oncogene. 1997 Sep 4;15(10):1179-89 PMID: 9294611
  51. p53 is phosphorylated in vitro and in vivo by the delta and epsilon isoforms of casein kinase 1 and enhances the level of casein kinase 1 delta in response to topoisomerase-directed drugs.
    Oncogene. 1997 Oct 2;15(14):1727-36 PMID: 9349507
  52. DNA damage-induced phosphorylation of p53 alleviates inhibition by MDM2.
    Cell. 1997 Oct 31;91(3):325-34 PMID: 9363941
  53. Rad53-dependent phosphorylation of Swi6 and down-regulation of CLN1 and CLN2 transcription occur in response to DNA damage in Saccharomyces cerevisiae.
    Genes Dev. 1997 Nov 15;11(22):3032-45 PMID: 9367985
  54. Atm-dependent interactions of a mammalian chk1 homolog with meiotic chromosomes.
    Curr Biol. 1997 Dec 1;7(12):977-86 PMID: 9382850
  55. DNA damage induces phosphorylation of the amino terminus of p53.
    Genes Dev. 1997 Dec 15;11(24):3471-81 PMID: 9407038
  56. The anaphase inhibitor of Saccharomyces cerevisiae Pds1p is a target of the DNA damage checkpoint pathway.
    Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14361-6 PMID: 9405617
  57. Nucleo-cytoplasmic shuttling of the hdm2 oncoprotein regulates the levels of the p53 protein via a pathway used by the human immunodeficiency virus rev protein.
    EMBO J. 1998 Jan 15;17(2):554-64 PMID: 9430646
  58. S-phase-specific activation of Cds1 kinase defines a subpathway of the checkpoint response in Schizosaccharomyces pombe.
    Genes Dev. 1998 Feb 1;12(3):382-95 PMID: 9450932
  59. Functional activation of p53 via phosphorylation following DNA damage by UV but not gamma radiation.
    Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2834-7 PMID: 9501176
  60. Replication checkpoint enforced by kinases Cds1 and Chk1.
    Science. 1998 May 8;280(5365):909-12 PMID: 9572736
  61. Absence of a radiation-induced first-cycle G1-S arrest in p53+ human tumor cells synchronized by mitotic selection.
    Cancer Res. 1998 May 1;58(9):2036-41 PMID: 9581850
  62. Identification of an additional negative regulatory region for p53 sequence-specific DNA binding.
    Proc Natl Acad Sci U S A. 1998 May 26;95(11):6079-84 PMID: 9600920
  63. Ultraviolet radiation, but not gamma radiation or etoposide-induced DNA damage, results in the phosphorylation of the murine p53 protein at serine-389.
    Proc Natl Acad Sci U S A. 1998 May 26;95(11):6399-402 PMID: 9600977
  64. ATM-dependent activation of p53 involves dephosphorylation and association with 14-3-3 proteins.
    Nat Genet. 1998 Jun;19(2):175-8 PMID: 9620776
  65. Enhanced phosphorylation of p53 by ATM in response to DNA damage.
    Science. 1998 Sep 11;281(5383):1674-7 PMID: 9733514
  66. Activation of the ATM kinase by ionizing radiation and phosphorylation of p53.
    Science. 1998 Sep 11;281(5383):1677-9 PMID: 9733515
  67. DNA damage and checkpoint pathways: molecular anatomy and interactions with repair.
    Cell. 1998 Sep 4;94(5):555-8 PMID: 9741620
  68. DNA damage activates p53 through a phosphorylation-acetylation cascade.
    Genes Dev. 1998 Sep 15;12(18):2831-41 PMID: 9744860
  69. DNA damage triggers DRB-resistant phosphorylation of human p53 at the CK2 site.
    Oncogene. 1998 Aug 27;17(8):1045-52 PMID: 9747884
  70. Replication checkpoint requires phosphorylation of the phosphatase Cdc25 by Cds1 or Chk1.
    Nature. 1998 Oct 1;395(6701):507-10 PMID: 9774107
  71. Diminished capacity for p53 in mediating a radiation-induced G1 arrest in established human tumor cell lines.
    Oncogene. 1995 Nov 2;11(9):1885-92 PMID: 7478618
  72. rad-dependent response of the chk1-encoded protein kinase at the DNA damage checkpoint.
    Science. 1996 Jan 19;271(5247):353-6 PMID: 8553071
  73. Regulation of RAD53 by the ATM-like kinases MEC1 and TEL1 in yeast cell cycle checkpoint pathways.
    Science. 1996 Jan 19;271(5247):357-60 PMID: 8553072
  74. Spk1/Rad53 is regulated by Mec1-dependent protein phosphorylation in DNA replication and damage checkpoint pathways.
    Genes Dev. 1996 Feb 15;10(4):395-406 PMID: 8600024
  75. cDNA cloning and gene mapping of a candidate human cell cycle checkpoint protein.
    Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2850-5 PMID: 8610130
  76. A reversible, p53-dependent G0/G1 cell cycle arrest induced by ribonucleotide depletion in the absence of detectable DNA damage.
    Genes Dev. 1996 Apr 15;10(8):934-47 PMID: 8608941
  77. A functionally inactive p53 protein in teratocarcinoma cells is activated by either DNA damage or cellular differentiation.
    Nat Med. 1996 Jul;2(7):804-10 PMID: 8673928
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
Published
2000-02-01
Pages
278-88
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316357
Subset
IM
Grants
NCI NIH HHS · CA09171 · United States
NCI NIH HHS · P01 CA025874 · United States
NCI NIH HHS · CA76367 · United States
NCI NIH HHS · R01 CA076367 · United States
NCI NIH HHS · T32 CA009171 · United States
NCI NIH HHS · CA25874 · United States
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2