Possible involvement of RecQL4 in the repair of double-strand DNA breaks in Xenopus egg extracts

Yuji Kumata, Shusuke Tada, Yumie Yamanada, Takashi Tsuyama, Takayuki Kobayashi, Yu Peng Dong, Kyoko Ikegami, Hiromu Murofushi, Masayuki Seki, Takemi Enomoto

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

Mutations in RecQL4 are a causative factor in Rothmund-Thomson syndrome, a human autosomal recessive disorder characterized by premature aging. To study the role of RecQL4, we employed a cell-free experimental system consisting of Xenopus egg extracts. RecQL4 loading onto chromatin was observed regardless of the presence or absence of EcoRI. However, in the absence of EcoRI, RecQL4 loading was suppressed by geminin, an inhibitor of pre-replicative complex formation, while in the presence of EcoRI, it was not affected. These results suggest that under the former condition, RecQL4-loading depended on DNA replication, while under the latter, the interaction occurred in response to double-stranded DNA breaks (DSBs) induced by EcoRI. DSB-induced RecQL4 loading depended on the function of the ataxia-telangiectasia mutated protein, DNA-dependent protein kinase (DNA-PK), and replication protein A, while there were only minor changes in DNA replication-associated RecQL4 loading upon suppression of these proteins. Furthermore, analyses using a chromatin-immunoprecipitation assay and quantification of γH2AX after induction of DSBs suggested that RecQL4 is loaded adjacent to Ku heterodimer-binding sites on damaged chromatin, and functions in the repair of DSBs.

Original languageEnglish
Pages (from-to)556-564
Number of pages9
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1773
Issue number4
DOIs
Publication statusPublished - 2007 Apr

Keywords

  • DNA-dependent protein kinase (DNA-PK)
  • Double-strand DNA break repair
  • Ku heterodimer
  • Non-homologous end-joining (NHEJ)
  • RecQL4

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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