DNA Damage Induces p53-dependent Down-regulation of hCHK1

Giovanna Damia, Yolanda Sanchez, Eugenio Erba, Massimo Broggini

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The levels of the human checkpoint gene hCHK1 were measured in human cancer cells growing in vitro after treatment with the DNA damaging agent cis-dichlorodi-ammine platinum(II) (DDP). Treatment of human cancer cell lines with DDP induced a decrease in the hCHK1 protein levels starting 6 h after treatment, with a further decline at 24 and 48 h. A similar decrease in the levels of hCHK1 was found at the mRNA level by using Northern blot analysis. By using isogenic cell systems in which p53 was disrupted either by transfection with HPV-E6 or by targeted homologous recombination, we found that the DNA damage-induced down-regulation of hCHK1 was only observable in wild type p53-expressing cells, with only a minor decline in the hCHK1 levels observable 48 h after treatment in cells with disrupted p53. Similarly, treatment of mutant p53-expressing human cancer cell lines with DDP did not result in changes in the levels of hCHK1. The p53-dependent down-regulation of hCHK1 is likely to be at transcriptional levels, as suggested by the lack of down-regulation of the hCHK1 when transfected under the control of a heterologous viral promoter. In addition, p53 is able to down-regulate the luciferase activity under the control of the 5′ flanking region of the hCHK1 gene. The data suggest a strict link between p53 and hCHK1 governing the activation and repression of the G2 checkpoint in which both proteins participate.

Original languageEnglish
Pages (from-to)10641-10645
Number of pages5
JournalJournal of Biological Chemistry
Volume276
Issue number14
DOIs
Publication statusPublished - Apr 6 2001

Fingerprint

DNA Damage
Down-Regulation
Cells
DNA
Genes
5' Flanking Region
Platinum
Luciferases
Cell Line
Neoplasms
Homologous Recombination
Proteins
Chemical activation
Northern Blotting
Transfection
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

DNA Damage Induces p53-dependent Down-regulation of hCHK1. / Damia, Giovanna; Sanchez, Yolanda; Erba, Eugenio; Broggini, Massimo.

In: Journal of Biological Chemistry, Vol. 276, No. 14, 06.04.2001, p. 10641-10645.

Research output: Contribution to journalArticle

Damia, Giovanna ; Sanchez, Yolanda ; Erba, Eugenio ; Broggini, Massimo. / DNA Damage Induces p53-dependent Down-regulation of hCHK1. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 14. pp. 10641-10645.
@article{e79c7e1a64844ae8841746f945aa53e0,
title = "DNA Damage Induces p53-dependent Down-regulation of hCHK1",
abstract = "The levels of the human checkpoint gene hCHK1 were measured in human cancer cells growing in vitro after treatment with the DNA damaging agent cis-dichlorodi-ammine platinum(II) (DDP). Treatment of human cancer cell lines with DDP induced a decrease in the hCHK1 protein levels starting 6 h after treatment, with a further decline at 24 and 48 h. A similar decrease in the levels of hCHK1 was found at the mRNA level by using Northern blot analysis. By using isogenic cell systems in which p53 was disrupted either by transfection with HPV-E6 or by targeted homologous recombination, we found that the DNA damage-induced down-regulation of hCHK1 was only observable in wild type p53-expressing cells, with only a minor decline in the hCHK1 levels observable 48 h after treatment in cells with disrupted p53. Similarly, treatment of mutant p53-expressing human cancer cell lines with DDP did not result in changes in the levels of hCHK1. The p53-dependent down-regulation of hCHK1 is likely to be at transcriptional levels, as suggested by the lack of down-regulation of the hCHK1 when transfected under the control of a heterologous viral promoter. In addition, p53 is able to down-regulate the luciferase activity under the control of the 5′ flanking region of the hCHK1 gene. The data suggest a strict link between p53 and hCHK1 governing the activation and repression of the G2 checkpoint in which both proteins participate.",
author = "Giovanna Damia and Yolanda Sanchez and Eugenio Erba and Massimo Broggini",
year = "2001",
month = "4",
day = "6",
doi = "10.1074/jbc.M007178200",
language = "English",
volume = "276",
pages = "10641--10645",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "14",

}

TY - JOUR

T1 - DNA Damage Induces p53-dependent Down-regulation of hCHK1

AU - Damia, Giovanna

AU - Sanchez, Yolanda

AU - Erba, Eugenio

AU - Broggini, Massimo

PY - 2001/4/6

Y1 - 2001/4/6

N2 - The levels of the human checkpoint gene hCHK1 were measured in human cancer cells growing in vitro after treatment with the DNA damaging agent cis-dichlorodi-ammine platinum(II) (DDP). Treatment of human cancer cell lines with DDP induced a decrease in the hCHK1 protein levels starting 6 h after treatment, with a further decline at 24 and 48 h. A similar decrease in the levels of hCHK1 was found at the mRNA level by using Northern blot analysis. By using isogenic cell systems in which p53 was disrupted either by transfection with HPV-E6 or by targeted homologous recombination, we found that the DNA damage-induced down-regulation of hCHK1 was only observable in wild type p53-expressing cells, with only a minor decline in the hCHK1 levels observable 48 h after treatment in cells with disrupted p53. Similarly, treatment of mutant p53-expressing human cancer cell lines with DDP did not result in changes in the levels of hCHK1. The p53-dependent down-regulation of hCHK1 is likely to be at transcriptional levels, as suggested by the lack of down-regulation of the hCHK1 when transfected under the control of a heterologous viral promoter. In addition, p53 is able to down-regulate the luciferase activity under the control of the 5′ flanking region of the hCHK1 gene. The data suggest a strict link between p53 and hCHK1 governing the activation and repression of the G2 checkpoint in which both proteins participate.

AB - The levels of the human checkpoint gene hCHK1 were measured in human cancer cells growing in vitro after treatment with the DNA damaging agent cis-dichlorodi-ammine platinum(II) (DDP). Treatment of human cancer cell lines with DDP induced a decrease in the hCHK1 protein levels starting 6 h after treatment, with a further decline at 24 and 48 h. A similar decrease in the levels of hCHK1 was found at the mRNA level by using Northern blot analysis. By using isogenic cell systems in which p53 was disrupted either by transfection with HPV-E6 or by targeted homologous recombination, we found that the DNA damage-induced down-regulation of hCHK1 was only observable in wild type p53-expressing cells, with only a minor decline in the hCHK1 levels observable 48 h after treatment in cells with disrupted p53. Similarly, treatment of mutant p53-expressing human cancer cell lines with DDP did not result in changes in the levels of hCHK1. The p53-dependent down-regulation of hCHK1 is likely to be at transcriptional levels, as suggested by the lack of down-regulation of the hCHK1 when transfected under the control of a heterologous viral promoter. In addition, p53 is able to down-regulate the luciferase activity under the control of the 5′ flanking region of the hCHK1 gene. The data suggest a strict link between p53 and hCHK1 governing the activation and repression of the G2 checkpoint in which both proteins participate.

UR - http://www.scopus.com/inward/record.url?scp=0035815627&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035815627&partnerID=8YFLogxK

U2 - 10.1074/jbc.M007178200

DO - 10.1074/jbc.M007178200

M3 - Article

C2 - 11152453

AN - SCOPUS:0035815627

VL - 276

SP - 10641

EP - 10645

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 14

ER -