Identification and characterization of MUS81 point mutations that abolish interaction with the SLX4 scaffold protein

Nidhi Nair, Dennis Castor, Thomas Macartney, John Rouse (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    13 Citations (Scopus)
    94 Downloads (Pure)

    Abstract

    MUS81-EME1 is a conserved structure-selective endonuclease with a preference for branched DNA substrates in vitro that correspond to intermediates of DNA repair. Cells lacking MUS81 or EME1 show defects in the repair of DNA interstrand crosslinks (ICL) resulting in hypersensitivity to agents such as mitomycin C. In metazoans, a proportion of cellular MUS81-EME1 binds the SLX4 scaffold protein, which is itself instrumental for ICL repair. It was previously reported that mutations in SLX4 that abolished interaction with MUS81 affected ICL repair in human cells but not in murine cells. In this study we looked the other way around by pinpointing amino acid residues in MUS81 that when mutated abolish the interaction with SLX4. These mutations fully rescued the mitomycin C hypersensitivity of MUS81 knockout murine cells, but they were unable to rescue the sensitivity of two different human cell lines defective in MUS81. These data support an SLX4-dependent role for MUS81 in the repair, but not the induction of ICL-induced double-strand breaks. This study sheds light on the extent to which MUS81 function in ICL repair requires interaction with SLX4.
    Original languageEnglish
    Pages (from-to)131-137
    Number of pages7
    JournalDNA Repair
    Volume24
    DOIs
    Publication statusPublished - Dec 2014

    Fingerprint

    Point Mutation
    Scaffolds
    Repair
    Mitomycin
    DNA Repair
    Hypersensitivity
    Proteins
    Mutation
    Endonucleases
    DNA
    Cells
    Amino Acids
    Cell Line
    Defects
    Substrates

    Cite this

    @article{01eeaa4e634c484eb20ffd8b37e9c5c8,
    title = "Identification and characterization of MUS81 point mutations that abolish interaction with the SLX4 scaffold protein",
    abstract = "MUS81-EME1 is a conserved structure-selective endonuclease with a preference for branched DNA substrates in vitro that correspond to intermediates of DNA repair. Cells lacking MUS81 or EME1 show defects in the repair of DNA interstrand crosslinks (ICL) resulting in hypersensitivity to agents such as mitomycin C. In metazoans, a proportion of cellular MUS81-EME1 binds the SLX4 scaffold protein, which is itself instrumental for ICL repair. It was previously reported that mutations in SLX4 that abolished interaction with MUS81 affected ICL repair in human cells but not in murine cells. In this study we looked the other way around by pinpointing amino acid residues in MUS81 that when mutated abolish the interaction with SLX4. These mutations fully rescued the mitomycin C hypersensitivity of MUS81 knockout murine cells, but they were unable to rescue the sensitivity of two different human cell lines defective in MUS81. These data support an SLX4-dependent role for MUS81 in the repair, but not the induction of ICL-induced double-strand breaks. This study sheds light on the extent to which MUS81 function in ICL repair requires interaction with SLX4.",
    author = "Nidhi Nair and Dennis Castor and Thomas Macartney and John Rouse",
    note = "Copyright {\circledC} 2014 The Authors. Published by Elsevier B.V. All rights reserved.",
    year = "2014",
    month = "12",
    doi = "10.1016/j.dnarep.2014.08.004",
    language = "English",
    volume = "24",
    pages = "131--137",
    journal = "DNA Repair",
    issn = "1568-7864",
    publisher = "Elsevier",

    }

    Identification and characterization of MUS81 point mutations that abolish interaction with the SLX4 scaffold protein. / Nair, Nidhi; Castor, Dennis; Macartney, Thomas; Rouse, John (Lead / Corresponding author).

    In: DNA Repair, Vol. 24, 12.2014, p. 131-137.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Identification and characterization of MUS81 point mutations that abolish interaction with the SLX4 scaffold protein

    AU - Nair, Nidhi

    AU - Castor, Dennis

    AU - Macartney, Thomas

    AU - Rouse, John

    N1 - Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

    PY - 2014/12

    Y1 - 2014/12

    N2 - MUS81-EME1 is a conserved structure-selective endonuclease with a preference for branched DNA substrates in vitro that correspond to intermediates of DNA repair. Cells lacking MUS81 or EME1 show defects in the repair of DNA interstrand crosslinks (ICL) resulting in hypersensitivity to agents such as mitomycin C. In metazoans, a proportion of cellular MUS81-EME1 binds the SLX4 scaffold protein, which is itself instrumental for ICL repair. It was previously reported that mutations in SLX4 that abolished interaction with MUS81 affected ICL repair in human cells but not in murine cells. In this study we looked the other way around by pinpointing amino acid residues in MUS81 that when mutated abolish the interaction with SLX4. These mutations fully rescued the mitomycin C hypersensitivity of MUS81 knockout murine cells, but they were unable to rescue the sensitivity of two different human cell lines defective in MUS81. These data support an SLX4-dependent role for MUS81 in the repair, but not the induction of ICL-induced double-strand breaks. This study sheds light on the extent to which MUS81 function in ICL repair requires interaction with SLX4.

    AB - MUS81-EME1 is a conserved structure-selective endonuclease with a preference for branched DNA substrates in vitro that correspond to intermediates of DNA repair. Cells lacking MUS81 or EME1 show defects in the repair of DNA interstrand crosslinks (ICL) resulting in hypersensitivity to agents such as mitomycin C. In metazoans, a proportion of cellular MUS81-EME1 binds the SLX4 scaffold protein, which is itself instrumental for ICL repair. It was previously reported that mutations in SLX4 that abolished interaction with MUS81 affected ICL repair in human cells but not in murine cells. In this study we looked the other way around by pinpointing amino acid residues in MUS81 that when mutated abolish the interaction with SLX4. These mutations fully rescued the mitomycin C hypersensitivity of MUS81 knockout murine cells, but they were unable to rescue the sensitivity of two different human cell lines defective in MUS81. These data support an SLX4-dependent role for MUS81 in the repair, but not the induction of ICL-induced double-strand breaks. This study sheds light on the extent to which MUS81 function in ICL repair requires interaction with SLX4.

    U2 - 10.1016/j.dnarep.2014.08.004

    DO - 10.1016/j.dnarep.2014.08.004

    M3 - Article

    C2 - 25224045

    VL - 24

    SP - 131

    EP - 137

    JO - DNA Repair

    JF - DNA Repair

    SN - 1568-7864

    ER -