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

    Fingerprint

    Replication Protein A
    DNA
    Fanconi Anemia
    Endonucleases
    Repair
    Fanconi Anemia Complementation Group Proteins
    Single-Stranded DNA
    DNA Replication
    Recombinant Proteins

    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

    Cite this

    Human FAN1 promotes strand incision in 5′-flapped DNA complexed with RPA. / Takahashi, Daisuke; Sato, Koichi; Hirayama, Emiko; Takata, Minoru; Kurumizaka, Hitoshi.

    In: Journal of Biochemistry, Vol. 158, No. 3, 2015, p. 263-270.

    Research output: Contribution to journalArticle

    Takahashi, Daisuke ; Sato, Koichi ; Hirayama, Emiko ; Takata, Minoru ; Kurumizaka, Hitoshi. / Human FAN1 promotes strand incision in 5′-flapped DNA complexed with RPA. In: Journal of Biochemistry. 2015 ; Vol. 158, No. 3. pp. 263-270.
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    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.",
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    AU - Takahashi, Daisuke

    AU - Sato, Koichi

    AU - Hirayama, Emiko

    AU - Takata, Minoru

    AU - Kurumizaka, Hitoshi

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    N2 - 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.

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