ATR-Chk1 signaling pathway and homologous recombinational repair protect cells from 5-fluorouracil cytotoxicity

Yoshihiko Fujinaka, Kazuaki Matsuoka, Makoto Iimori, Munkhbold Tuul, Ryo Sakasai, Keiji Yoshinaga, Hiroshi Saeki, Masaru Morita, Yoshihiro Kakeji, David A. Gillespie, Ken-ichi Yamamoto, Minoru Takata, Hiroyuki Kitao, Yoshihiko Maehara

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

5-Fluorouracil (5-FU) has long been a mainstay antimetabolite chemotherapeutic drug for the treatment of major solid tumors, particularly colorectal cancer. 5-FU is processed intracellularly to yield active metabolites that compromise RNA and DNA metabolism. However, the mechanisms responsible for its cytotoxicity are not fully understood. From the phenotypic analysis of mutant chicken B lymphoma DT40 cells, we found that homologous recombinational repair (HRR), involving Rad54 and BRCA2, and the ATR-Chk1 signaling pathway, involving Rad9 and Rad17, significantly contribute to 5-FU tolerance. 5-FU induced γH2AX nuclear foci, which were colocalized with the key HRR factor Rad51, but not with DNA double-strand breaks (DSBs), in a dose-dependent manner as cells accumulated in the S phase. Inhibition of Chk1 kinase by UCN-01 increased 5-FU-induced γH2AX and enhanced 5-FU cytotoxicity not only in wild-type cells but also in Rad54- or BRCA2-deficient cells, suggesting that HRR and Chk1 kinase have non-overlapping roles in 5-FU tolerance. 5-FU-induced Chk1 phosphorylation was significantly impaired in Rad9- or Rad17-deficient cells, and severe γH2AX nuclear foci and DSBs were formed, which was followed by apoptosis. Finally, inhibition of Chk1 kinase by UCN-01 increased 5-FU-induced γH2AX nuclear foci and enhanced 5-FU cytotoxicity in Rad9- or Rad17-deficient cells. These results suggest that Rad9- and Rad17-independent activation of the ATR-Chk1 signaling pathway also significantly contributes to 5-FU tolerance.

Original languageEnglish
Pages (from-to)247-258
Number of pages12
JournalDNA Repair
Volume11
Issue number3
DOIs
Publication statusPublished - Mar 1 2012
Externally publishedYes

Fingerprint

Recombinational DNA Repair
Cytotoxicity
Fluorouracil
Repair
Phosphotransferases
Antimetabolites
Phosphorylation
Double-Stranded DNA Breaks
DNA
B-Cell Lymphoma
Metabolites
S Phase
Metabolism
Tumors
Colorectal Neoplasms
Chickens
Chemical activation

Keywords

  • 5-Fluorouracil
  • Chk1
  • Double-strand breaks
  • Homologous recombinational repair
  • Rad17
  • Rad9

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Fujinaka, Y., Matsuoka, K., Iimori, M., Tuul, M., Sakasai, R., Yoshinaga, K., ... Maehara, Y. (2012). ATR-Chk1 signaling pathway and homologous recombinational repair protect cells from 5-fluorouracil cytotoxicity. DNA Repair, 11(3), 247-258. https://doi.org/10.1016/j.dnarep.2011.11.005

ATR-Chk1 signaling pathway and homologous recombinational repair protect cells from 5-fluorouracil cytotoxicity. / Fujinaka, Yoshihiko; Matsuoka, Kazuaki; Iimori, Makoto; Tuul, Munkhbold; Sakasai, Ryo; Yoshinaga, Keiji; Saeki, Hiroshi; Morita, Masaru; Kakeji, Yoshihiro; Gillespie, David A.; Yamamoto, Ken-ichi; Takata, Minoru; Kitao, Hiroyuki; Maehara, Yoshihiko.

In: DNA Repair, Vol. 11, No. 3, 01.03.2012, p. 247-258.

Research output: Contribution to journalArticle

Fujinaka, Y, Matsuoka, K, Iimori, M, Tuul, M, Sakasai, R, Yoshinaga, K, Saeki, H, Morita, M, Kakeji, Y, Gillespie, DA, Yamamoto, K, Takata, M, Kitao, H & Maehara, Y 2012, 'ATR-Chk1 signaling pathway and homologous recombinational repair protect cells from 5-fluorouracil cytotoxicity', DNA Repair, vol. 11, no. 3, pp. 247-258. https://doi.org/10.1016/j.dnarep.2011.11.005
Fujinaka, Yoshihiko ; Matsuoka, Kazuaki ; Iimori, Makoto ; Tuul, Munkhbold ; Sakasai, Ryo ; Yoshinaga, Keiji ; Saeki, Hiroshi ; Morita, Masaru ; Kakeji, Yoshihiro ; Gillespie, David A. ; Yamamoto, Ken-ichi ; Takata, Minoru ; Kitao, Hiroyuki ; Maehara, Yoshihiko. / ATR-Chk1 signaling pathway and homologous recombinational repair protect cells from 5-fluorouracil cytotoxicity. In: DNA Repair. 2012 ; Vol. 11, No. 3. pp. 247-258.
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