Yeast RAD7-RAD16 complex, specific for the nucleotide excision repair of the nontranscribed DNA strand, is an ATP-dependent DNA damage sensor

Sami N. Guzder, Patrick Sung, Louise Prakash, Satya Prakash

    Research output: Contribution to journalArticle

    67 Citations (Scopus)

    Abstract

    In eukaryotes, nucleotide excision repair of ultraviolet light-damaged DNA is a highly intricate process that requires a large number of evolutionarily conserved protein factors. Genetic studies in the yeast Saccharomyces cerevisiae have indicated a specific role of the RAD7 and RAD16 genes in the repair of transcriptionally inactive DNA. Here we show that the RAD7- and RAD16-encoded products exist as a complex of 1:1 stoichiometry, exhibiting an apparent dissociation constant (K(d)) of <4 x 10-10 M. The RAD7-RAd16 complex has been purified to near homogeneity in this study and is shown to bind, in an ATP-dependent manner and with high specificity, to DNA damaged by ultraviolet light. Importantly, inclusion of the RAD7-RAD16 complex in the in vitro nucleotide excision repair system that consists entirely of purified components results in a marked stimulation of damage specific incision. Thus, Rad7-Rad16 complex is the ATP-dependent DNA damage sensor that specifically functions with the ensemble of nucleotide excision repair factor (NEF) 1, NEF2, NEF3, and replication protein A in the repair of transcriptionally inactive DNA. We name this novel complex of Rad7 and Rad16 proteins NEF4.

    Original languageEnglish
    Pages (from-to)21665-21668
    Number of pages4
    JournalJournal of Biological Chemistry
    Volume272
    Issue number35
    DOIs
    StatePublished - Aug 29 1997

    Fingerprint

    DNA Repair
    Yeast
    DNA Damage
    Repair
    Nucleotides
    Adenosine Triphosphate
    Yeasts
    DNA
    Sensors
    Ultraviolet Rays
    Replication Protein A
    Eukaryota
    Names
    Saccharomyces cerevisiae
    Proteins
    Stoichiometry
    Genes

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Yeast RAD7-RAD16 complex, specific for the nucleotide excision repair of the nontranscribed DNA strand, is an ATP-dependent DNA damage sensor. / Guzder, Sami N.; Sung, Patrick; Prakash, Louise; Prakash, Satya.

    In: Journal of Biological Chemistry, Vol. 272, No. 35, 29.08.1997, p. 21665-21668.

    Research output: Contribution to journalArticle

    @article{612761768f6a459c9f0ea02ab67d8e8c,
    title = "Yeast RAD7-RAD16 complex, specific for the nucleotide excision repair of the nontranscribed DNA strand, is an ATP-dependent DNA damage sensor",
    abstract = "In eukaryotes, nucleotide excision repair of ultraviolet light-damaged DNA is a highly intricate process that requires a large number of evolutionarily conserved protein factors. Genetic studies in the yeast Saccharomyces cerevisiae have indicated a specific role of the RAD7 and RAD16 genes in the repair of transcriptionally inactive DNA. Here we show that the RAD7- and RAD16-encoded products exist as a complex of 1:1 stoichiometry, exhibiting an apparent dissociation constant (K(d)) of <4 x 10-10 M. The RAD7-RAd16 complex has been purified to near homogeneity in this study and is shown to bind, in an ATP-dependent manner and with high specificity, to DNA damaged by ultraviolet light. Importantly, inclusion of the RAD7-RAD16 complex in the in vitro nucleotide excision repair system that consists entirely of purified components results in a marked stimulation of damage specific incision. Thus, Rad7-Rad16 complex is the ATP-dependent DNA damage sensor that specifically functions with the ensemble of nucleotide excision repair factor (NEF) 1, NEF2, NEF3, and replication protein A in the repair of transcriptionally inactive DNA. We name this novel complex of Rad7 and Rad16 proteins NEF4.",
    author = "Guzder, {Sami N.} and Patrick Sung and Louise Prakash and Satya Prakash",
    year = "1997",
    month = "8",
    day = "29",
    doi = "10.1074/jbc.272.35.21665",
    language = "English",
    volume = "272",
    pages = "21665--21668",
    journal = "Journal of Biological Chemistry",
    issn = "0021-9258",
    publisher = "American Society for Biochemistry and Molecular Biology Inc.",
    number = "35",

    }

    TY - JOUR

    T1 - Yeast RAD7-RAD16 complex, specific for the nucleotide excision repair of the nontranscribed DNA strand, is an ATP-dependent DNA damage sensor

    AU - Guzder, Sami N.

    AU - Sung, Patrick

    AU - Prakash, Louise

    AU - Prakash, Satya

    PY - 1997/8/29

    Y1 - 1997/8/29

    N2 - In eukaryotes, nucleotide excision repair of ultraviolet light-damaged DNA is a highly intricate process that requires a large number of evolutionarily conserved protein factors. Genetic studies in the yeast Saccharomyces cerevisiae have indicated a specific role of the RAD7 and RAD16 genes in the repair of transcriptionally inactive DNA. Here we show that the RAD7- and RAD16-encoded products exist as a complex of 1:1 stoichiometry, exhibiting an apparent dissociation constant (K(d)) of <4 x 10-10 M. The RAD7-RAd16 complex has been purified to near homogeneity in this study and is shown to bind, in an ATP-dependent manner and with high specificity, to DNA damaged by ultraviolet light. Importantly, inclusion of the RAD7-RAD16 complex in the in vitro nucleotide excision repair system that consists entirely of purified components results in a marked stimulation of damage specific incision. Thus, Rad7-Rad16 complex is the ATP-dependent DNA damage sensor that specifically functions with the ensemble of nucleotide excision repair factor (NEF) 1, NEF2, NEF3, and replication protein A in the repair of transcriptionally inactive DNA. We name this novel complex of Rad7 and Rad16 proteins NEF4.

    AB - In eukaryotes, nucleotide excision repair of ultraviolet light-damaged DNA is a highly intricate process that requires a large number of evolutionarily conserved protein factors. Genetic studies in the yeast Saccharomyces cerevisiae have indicated a specific role of the RAD7 and RAD16 genes in the repair of transcriptionally inactive DNA. Here we show that the RAD7- and RAD16-encoded products exist as a complex of 1:1 stoichiometry, exhibiting an apparent dissociation constant (K(d)) of <4 x 10-10 M. The RAD7-RAd16 complex has been purified to near homogeneity in this study and is shown to bind, in an ATP-dependent manner and with high specificity, to DNA damaged by ultraviolet light. Importantly, inclusion of the RAD7-RAD16 complex in the in vitro nucleotide excision repair system that consists entirely of purified components results in a marked stimulation of damage specific incision. Thus, Rad7-Rad16 complex is the ATP-dependent DNA damage sensor that specifically functions with the ensemble of nucleotide excision repair factor (NEF) 1, NEF2, NEF3, and replication protein A in the repair of transcriptionally inactive DNA. We name this novel complex of Rad7 and Rad16 proteins NEF4.

    UR - http://www.scopus.com/inward/record.url?scp=0030856090&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0030856090&partnerID=8YFLogxK

    U2 - 10.1074/jbc.272.35.21665

    DO - 10.1074/jbc.272.35.21665

    M3 - Article

    VL - 272

    SP - 21665

    EP - 21668

    JO - Journal of Biological Chemistry

    JF - Journal of Biological Chemistry

    SN - 0021-9258

    IS - 35

    ER -