Human FAN1 promotes strand incision in 5′-flapped DNA complexed with RPA

Daisuke Takahashi, Koichi Sato, Emiko Hirayama, Minoru Takata, Hitoshi Kurumizaka

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Fanconi anaemia (FA) is a human infantile recessive disorder. Seventeen FA causal proteins cooperatively function in the DNA interstrand crosslink (ICL) repair pathway. Dual DNA strand incisions around the crosslink are critical steps in ICL repair. FA-associated nuclease 1 (FAN1) is a DNA structure-specific endonuclease that is considered to be involved in DNA incision at the stalled replication fork. Replication protein A (RPA) rapidly assembles on the single-stranded DNA region of the stalled fork. However, the effect of RPA on the FAN1-mediated DNA incision has not been determined. In this study, we purified human FAN1, as a bacterially expressed recombinant protein. FAN1 exhibited robust endonuclease activity with 5′-flapped DNA, which is formed at the stalled replication fork. We found that FAN1 efficiently promoted DNA incision at the proper site of RPA-coated 50-flapped DNA. Therefore, FAN1 possesses the ability to promote the ICL repair of 50-flapped DNA covered by RPA.

Original languageEnglish
Pages (from-to)263-270
Number of pages8
JournalJournal of biochemistry
Volume158
Issue number3
DOIs
Publication statusPublished - 2015

Keywords

  • DNA interstrand crosslinking repair
  • Fanconi anaemia
  • Fanconi anaemia-associated nuclease 1 (FAN1)
  • Replication protein A (RPA)
  • Structure-specific endonuclease

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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