Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint

T. Shimura, M. Toyoshima, S. K. Adiga, T. Kunoh, H. Nagai, N. Shimizu, M. Inoue, O. Niwa

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The S-phase DNA damage checkpoint is activated by DNA damage to delay DNA synthesis allowing time to resolve the replication block. We previously discovered the p53-dependent S-phase DNA damage checkpoint in mouse zygotes fertilized with irradiated sperm. Here, we report that the same p53 dependency holds in mouse embryonic fibroblasts (MEFs) at low doses of irradiation. DNA synthesis in p53 wild-type (WT) MEFs was suppressed in a biphasic manner in which a sharp decrease below 2.5 Gy was followed by a more moderate decrease up to 10 Gy. In contrast, p53-/- MEFs exhibited radioresistant DNA synthesis below 2.5 Gy whereas the cells retained the moderate suppression above 5 Gy. DNA fiber analysis revealed that 1 Gy irradiation suppressed replication fork progression in p53 WT MEFs, but not in p53-/- MEFs. Proliferating cell nuclear antigen (PCNA), clamp loader of DNA polymerase, was phosphorylated in WT MEFs after 1 Gy irradiation and redistributed to form foci in the nuclei. In contrast, PCNA was not phosphorylated and dissociated from chromatin in 1 Gy-irradiated p53-/- MEFs. These results demonstrate that the novel low-dose-specific p53-dependent S-phase DNA damage checkpoint is likely to regulate the replication fork movement through phosphorylation of PCNA.

Original languageEnglish
Pages (from-to)5921-5932
Number of pages12
JournalOncogene
Volume25
Issue number44
DOIs
Publication statusPublished - Sep 28 2006
Externally publishedYes

Fingerprint

S Phase
DNA Damage
Fibroblasts
Proliferating Cell Nuclear Antigen
DNA
Zygote
DNA-Directed DNA Polymerase
Chromatin
Spermatozoa
Phosphorylation

Keywords

  • ATM
  • p53
  • PCNA
  • Replication fork progression
  • S-phase DNA damage checkpoint

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Shimura, T., Toyoshima, M., Adiga, S. K., Kunoh, T., Nagai, H., Shimizu, N., ... Niwa, O. (2006). Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint. Oncogene, 25(44), 5921-5932. https://doi.org/10.1038/sj.onc.1209624

Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint. / Shimura, T.; Toyoshima, M.; Adiga, S. K.; Kunoh, T.; Nagai, H.; Shimizu, N.; Inoue, M.; Niwa, O.

In: Oncogene, Vol. 25, No. 44, 28.09.2006, p. 5921-5932.

Research output: Contribution to journalArticle

Shimura, T, Toyoshima, M, Adiga, SK, Kunoh, T, Nagai, H, Shimizu, N, Inoue, M & Niwa, O 2006, 'Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint', Oncogene, vol. 25, no. 44, pp. 5921-5932. https://doi.org/10.1038/sj.onc.1209624
Shimura T, Toyoshima M, Adiga SK, Kunoh T, Nagai H, Shimizu N et al. Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint. Oncogene. 2006 Sep 28;25(44):5921-5932. https://doi.org/10.1038/sj.onc.1209624
Shimura, T. ; Toyoshima, M. ; Adiga, S. K. ; Kunoh, T. ; Nagai, H. ; Shimizu, N. ; Inoue, M. ; Niwa, O. / Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint. In: Oncogene. 2006 ; Vol. 25, No. 44. pp. 5921-5932.
@article{14df2d033ccb48a885abff9dc161bd2c,
title = "Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint",
abstract = "The S-phase DNA damage checkpoint is activated by DNA damage to delay DNA synthesis allowing time to resolve the replication block. We previously discovered the p53-dependent S-phase DNA damage checkpoint in mouse zygotes fertilized with irradiated sperm. Here, we report that the same p53 dependency holds in mouse embryonic fibroblasts (MEFs) at low doses of irradiation. DNA synthesis in p53 wild-type (WT) MEFs was suppressed in a biphasic manner in which a sharp decrease below 2.5 Gy was followed by a more moderate decrease up to 10 Gy. In contrast, p53-/- MEFs exhibited radioresistant DNA synthesis below 2.5 Gy whereas the cells retained the moderate suppression above 5 Gy. DNA fiber analysis revealed that 1 Gy irradiation suppressed replication fork progression in p53 WT MEFs, but not in p53-/- MEFs. Proliferating cell nuclear antigen (PCNA), clamp loader of DNA polymerase, was phosphorylated in WT MEFs after 1 Gy irradiation and redistributed to form foci in the nuclei. In contrast, PCNA was not phosphorylated and dissociated from chromatin in 1 Gy-irradiated p53-/- MEFs. These results demonstrate that the novel low-dose-specific p53-dependent S-phase DNA damage checkpoint is likely to regulate the replication fork movement through phosphorylation of PCNA.",
keywords = "ATM, p53, PCNA, Replication fork progression, S-phase DNA damage checkpoint",
author = "T. Shimura and M. Toyoshima and Adiga, {S. K.} and T. Kunoh and H. Nagai and N. Shimizu and M. Inoue and O. Niwa",
year = "2006",
month = "9",
day = "28",
doi = "10.1038/sj.onc.1209624",
language = "English",
volume = "25",
pages = "5921--5932",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "44",

}

TY - JOUR

T1 - Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint

AU - Shimura, T.

AU - Toyoshima, M.

AU - Adiga, S. K.

AU - Kunoh, T.

AU - Nagai, H.

AU - Shimizu, N.

AU - Inoue, M.

AU - Niwa, O.

PY - 2006/9/28

Y1 - 2006/9/28

N2 - The S-phase DNA damage checkpoint is activated by DNA damage to delay DNA synthesis allowing time to resolve the replication block. We previously discovered the p53-dependent S-phase DNA damage checkpoint in mouse zygotes fertilized with irradiated sperm. Here, we report that the same p53 dependency holds in mouse embryonic fibroblasts (MEFs) at low doses of irradiation. DNA synthesis in p53 wild-type (WT) MEFs was suppressed in a biphasic manner in which a sharp decrease below 2.5 Gy was followed by a more moderate decrease up to 10 Gy. In contrast, p53-/- MEFs exhibited radioresistant DNA synthesis below 2.5 Gy whereas the cells retained the moderate suppression above 5 Gy. DNA fiber analysis revealed that 1 Gy irradiation suppressed replication fork progression in p53 WT MEFs, but not in p53-/- MEFs. Proliferating cell nuclear antigen (PCNA), clamp loader of DNA polymerase, was phosphorylated in WT MEFs after 1 Gy irradiation and redistributed to form foci in the nuclei. In contrast, PCNA was not phosphorylated and dissociated from chromatin in 1 Gy-irradiated p53-/- MEFs. These results demonstrate that the novel low-dose-specific p53-dependent S-phase DNA damage checkpoint is likely to regulate the replication fork movement through phosphorylation of PCNA.

AB - The S-phase DNA damage checkpoint is activated by DNA damage to delay DNA synthesis allowing time to resolve the replication block. We previously discovered the p53-dependent S-phase DNA damage checkpoint in mouse zygotes fertilized with irradiated sperm. Here, we report that the same p53 dependency holds in mouse embryonic fibroblasts (MEFs) at low doses of irradiation. DNA synthesis in p53 wild-type (WT) MEFs was suppressed in a biphasic manner in which a sharp decrease below 2.5 Gy was followed by a more moderate decrease up to 10 Gy. In contrast, p53-/- MEFs exhibited radioresistant DNA synthesis below 2.5 Gy whereas the cells retained the moderate suppression above 5 Gy. DNA fiber analysis revealed that 1 Gy irradiation suppressed replication fork progression in p53 WT MEFs, but not in p53-/- MEFs. Proliferating cell nuclear antigen (PCNA), clamp loader of DNA polymerase, was phosphorylated in WT MEFs after 1 Gy irradiation and redistributed to form foci in the nuclei. In contrast, PCNA was not phosphorylated and dissociated from chromatin in 1 Gy-irradiated p53-/- MEFs. These results demonstrate that the novel low-dose-specific p53-dependent S-phase DNA damage checkpoint is likely to regulate the replication fork movement through phosphorylation of PCNA.

KW - ATM

KW - p53

KW - PCNA

KW - Replication fork progression

KW - S-phase DNA damage checkpoint

UR - http://www.scopus.com/inward/record.url?scp=33749170308&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749170308&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1209624

DO - 10.1038/sj.onc.1209624

M3 - Article

C2 - 16682953

AN - SCOPUS:33749170308

VL - 25

SP - 5921

EP - 5932

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 44

ER -