Home LiteratureArticle Details
PMID: 10733583 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Posttranslational modifications of p53 in replicative senescence overlapping but distinct from those induced by DNA damage.

Molecular and cellular biology ·Vol. 20 ·No. 8 ·2000-04-00 ·Pages 2803-8

Webley K, Bond JA, Jones CJ, Blaydes JP, Craig A, Hupp T, Wynford-Thomas D

Abstract

Replicative senescence in human fibroblasts is absolutely dependent on the function of the phosphoprotein p53 and correlates with activation of p53-dependent transcription. However, no evidence for posttranslational modification of p53 in senescence has been presented, raising the possibility that changes in transcriptional activity result from upregulation of a coactivator. Using a series of antibodies with phosphorylation-sensitive epitopes, we now show that senescence is associated with major changes at putative regulatory sites in the N and C termini of p53 consistent with increased phosphorylation at serine-15, threonine-18, and serine-376 and decreased phosphorylation at serine-392. Ionizing and UV radiation generated overlapping but distinct profiles of response, with increased serine-15 phosphorylation being the only common change. These results support a direct role for p53 in signaling replicative senescence and are consistent with the generation by telomere erosion of a signal which shares some but not all of the features of DNA double-strand breaks.

MeSH Terms
Cell Line DNA Damage/radiation effects DNA Replication Fibroblasts Gene Expression Regulation Humans Protein Processing, Post-Translational Tumor Suppressor Protein p53/genetics,metabolism Ultraviolet Rays
Chemicals
Tumor Suppressor Protein p53
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Webley K
Cancer Research Campaign Laboratories, Department of Pathology, University of Wales College of Medicine, Cardiff CF14 4XN, United Kingdom.
Bond J A
Jones C J
Blaydes J P
Craig A
Hupp T
Wynford-Thomas D
References (58)
58 references, click to expand
  1. The candidate tumour suppressor p33ING1 cooperates with p53 in cell growth control.
    Nature. 1998 Jan 15;391(6664):295-8 PMID: 9440695
  2. Regulation of the sequence-specific DNA binding function of p53 by protein kinase C and protein phosphatases.
    J Biol Chem. 1995 Mar 10;270(10):5405-11 PMID: 7534296
  3. The genetic basis of human keratinocyte immortalisation in squamous cell carcinoma development: the role of telomerase reactivation.
    Eur J Cancer. 1997 Apr;33(5):727-34 PMID: 9282111
  4. Signaling to p53: breaking the MDM2-p53 circuit.
    Cell. 1998 Oct 2;95(1):5-8 PMID: 9778240
  5. Comparative study of the p53-mdm2 and p53-MDMX interfaces.
    Oncogene. 1999 Jan 7;18(1):189-99 PMID: 9926934
  6. Time, telomeres and tumours: is cellular senescence more than an anticancer mechanism?
    Trends Cell Biol. 1995 Aug;5(8):293-7 PMID: 14732086
  7. p53 mediates permanent arrest over multiple cell cycles in response to gamma-irradiation.
    Cancer Res. 1997 Mar 15;57(6):1171-9 PMID: 9067289
  8. 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
  9. DNA damage triggers a prolonged p53-dependent G1 arrest and long-term induction of Cip1 in normal human fibroblasts.
    Genes Dev. 1994 Nov 1;8(21):2540-51 PMID: 7958916
  10. S phase cell-cycle arrest following DNA damage is independent of the p53/p21(WAF1) signalling pathway.
    Oncogene. 1996 Mar 7;12(5):1077-82 PMID: 8649799
  11. DNA-dependent protein kinase acts upstream of p53 in response to DNA damage.
    Nature. 1998 Aug 13;394(6694):700-4 PMID: 9716137
  12. The p53-dependent G1 cell cycle checkpoint pathway and ataxia-telangiectasia.
    Cancer Res. 1994 Oct 1;54(19):5054-8 PMID: 7923116
  13. Mutant p53 rescues human diploid cells from senescence without inhibiting the induction of SDI1/WAF1.
    Cancer Res. 1995 Jun 1;55(11):2404-9 PMID: 7538902
  14. An immunochemical analysis of the human nuclear phosphoprotein p53. New monoclonal antibodies and epitope mapping using recombinant p53.
    J Immunol Methods. 1992 Jul 6;151(1-2):237-44 PMID: 1378473
  15. Telomerase prevents the accelerated cell ageing of Werner syndrome fibroblasts.
    Nat Genet. 2000 Jan;24(1):16-7 PMID: 10615119
  16. Cancer. p53, guardian of the genome.
    Nature. 1992 Jul 2;358(6381):15-6 PMID: 1614522
  17. Regulation of specific DNA binding by p53: evidence for a role for O-glycosylation and charged residues at the carboxy-terminus.
    Oncogene. 1996 Feb 15;12(4):921-30 PMID: 8632915
  18. Extension of the replicative life span of human diploid fibroblasts by inhibition of the p33ING1 candidate tumor suppressor.
    Mol Cell Biol. 1997 Apr;17(4):2014-9 PMID: 9121449
  19. Agents that cause DNA double strand breaks lead to p16INK4a enrichment and the premature senescence of normal fibroblasts.
    Oncogene. 1998 Mar 5;16(9):1113-23 PMID: 9528853
  20. DNA-dependent protein kinase is not required for the p53-dependent response to DNA damage.
    Nature. 1999 Jul 1;400(6739):81-3 PMID: 10403253
  21. ATM-dependent activation of p53 involves dephosphorylation and association with 14-3-3 proteins.
    Nat Genet. 1998 Jun;19(2):175-8 PMID: 9620776
  22. The ataxia-telangiectasia related protein ATR mediates DNA-dependent phosphorylation of p53.
    Oncogene. 1999 Jul 8;18(27):3989-95 PMID: 10435622
  23. The complexity of p53 modulation: emerging patterns from divergent signals.
    Genes Dev. 1998 Oct 1;12(19):2973-83 PMID: 9765199
  24. Small peptides activate the latent sequence-specific DNA binding function of p53.
    Cell. 1995 Oct 20;83(2):237-45 PMID: 7585941
  25. Dephosphorylation of p53 at Ser20 after cellular exposure to low levels of non-ionizing radiation.
    Oncogene. 1999 Nov 4;18(46):6305-12 PMID: 10597229
  26. DNA damage induces phosphorylation of the amino terminus of p53.
    Genes Dev. 1997 Dec 15;11(24):3471-81 PMID: 9407038
  27. THE LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS.
    Exp Cell Res. 1965 Mar;37:614-36 PMID: 14315085
  28. Evidence that transcriptional activation by p53 plays a direct role in the induction of cellular senescence.
    Oncogene. 1996 Nov 21;13(10):2097-104 PMID: 8950976
  29. Does telomere shortening drive selection for p53 mutation in human cancer?
    Mol Carcinog. 1995 Mar;12(3):119-23 PMID: 7893364
  30. Conformational changes in p53 analysed using new antibodies to the core DNA binding domain of the protein.
    Oncogene. 1995 Jan 19;10(2):389-93 PMID: 7530828
  31. Addition of fresh medium induces cell cycle and conformation changes in p53, a tumour suppressor protein.
    Oncogene. 1990 Nov;5(11):1683-90 PMID: 2176282
  32. Two critical hydrophobic amino acids in the N-terminal domain of the p53 protein are required for the gain of function phenotypes of human p53 mutants.
    Oncogene. 1995 Jun 15;10(12):2387-90 PMID: 7784087
  33. The biology of replicative senescence.
    Eur J Cancer. 1997 Apr;33(5):703-9 PMID: 9282108
  34. Allosteric activation of latent p53 tetramers.
    Curr Biol. 1994 Oct 1;4(10):865-75 PMID: 7850419
  35. Proliferative lifespan checkpoints: cell-type specificity and influence on tumour biology.
    Eur J Cancer. 1997 Apr;33(5):716-26 PMID: 9282110
  36. Characterisation of epitopes on human p53 using phage-displayed peptide libraries: insights into antibody-peptide interactions.
    J Mol Biol. 1995 Apr 21;248(1):58-78 PMID: 7537340
  37. Increased activity of p53 in senescing fibroblasts.
    Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8348-52 PMID: 7667293
  38. DNA damage activates p53 through a phosphorylation-acetylation cascade.
    Genes Dev. 1998 Sep 15;12(18):2831-41 PMID: 9744860
  39. A role for ATR in the DNA damage-induced phosphorylation of p53.
    Genes Dev. 1999 Jan 15;13(2):152-7 PMID: 9925639
  40. ATM associates with and phosphorylates p53: mapping the region of interaction.
    Nat Genet. 1998 Dec;20(4):398-400 PMID: 9843217
  41. Novel phosphorylation sites of human tumour suppressor protein p53 at Ser20 and Thr18 that disrupt the binding of mdm2 (mouse double minute 2) protein are modified in human cancers.
    Biochem J. 1999 Aug 15;342 ( Pt 1):133-41 PMID: 10432310
  42. DNA damage triggers DRB-resistant phosphorylation of human p53 at the CK2 site.
    Oncogene. 1998 Aug 27;17(8):1045-52 PMID: 9747884
  43. TRF2 protects human telomeres from end-to-end fusions.
    Cell. 1998 Feb 6;92(3):401-13 PMID: 9476899
  44. DNA damage-induced phosphorylation of p53 alleviates inhibition by MDM2.
    Cell. 1997 Oct 31;91(3):325-34 PMID: 9363941
  45. Extension of life-span by introduction of telomerase into normal human cells.
    Science. 1998 Jan 16;279(5349):349-52 PMID: 9454332
  46. 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
  47. Reinitiation of DNA synthesis and cell division in senescent human fibroblasts by microinjection of anti-p53 antibodies.
    Mol Cell Biol. 1998 Mar;18(3):1611-21 PMID: 9488478
  48. Regulation of the cryptic sequence-specific DNA-binding function of p53 by protein kinases.
    Cold Spring Harb Symp Quant Biol. 1994;59:195-206 PMID: 7587070
  49. Telomeres and telomerase in human cancer (review).
    Int J Oncol. 1995 Sep;7(3):423-32 PMID: 21552856
  50. The p53 pathway.
    J Pathol. 1999 Jan;187(1):112-26 PMID: 10341712
  51. Identification of novel mdm2 binding peptides by phage display.
    Oncogene. 1996 Nov 21;13(10):2141-7 PMID: 8950981
  52. Increased p53 protein associated with aging in human diploid fibroblasts.
    Exp Cell Res. 1995 Apr;217(2):336-45 PMID: 7698234
  53. p53-Dependent growth arrest and altered p53-immunoreactivity following metabolic labelling with 32P ortho-phosphate in human fibroblasts.
    Oncogene. 1999 Jun 24;18(25):3788-92 PMID: 10391688
  54. ATM-dependent telomere loss in aging human diploid fibroblasts and DNA damage lead to the post-translational activation of p53 protein involving poly(ADP-ribose) polymerase.
    EMBO J. 1997 Oct 1;16(19):6018-33 PMID: 9312059
  55. Protein interactions at the carboxyl terminus of p53 result in the induction of its in vitro transactivation potential.
    Oncogene. 1997 Jul 10;15(2):237-44 PMID: 9244359
  56. Activation of p53 sequence-specific DNA binding by acetylation of the p53 C-terminal domain.
    Cell. 1997 Aug 22;90(4):595-606 PMID: 9288740
  57. Escape from senescence in human diploid fibroblasts induced directly by mutant p53.
    Oncogene. 1994 Jul;9(7):1885-9 PMID: 8208534
  58. Cataloging altered gene expression in young and senescent cells using enhanced differential display.
    Nucleic Acids Res. 1995 Aug 25;23(16):3244-51 PMID: 7667101
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.20.8.2803-2808.2000
Published
2000-04-00
Pages
2803-8
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC85496
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.20.8.2803-2808.2000 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