Phosphorylation of Slx4 by Mec1 and Tel1 regulates the single-strand annealing mode of DNA repair in budding yeast

Sonja Flott, Constance Alabert, Geraldine W Toh, Rachel Toth, Neal Sugawara, David G Campbell, James E Haber, Philippe Pasero, John Rouse

    Research output: Contribution to journalArticle

    75 Citations (Scopus)

    Abstract

    Budding yeast (Saccharomyces cerevisiae) Slx4 is essential for cell viability in the absence of the Sgs1 helicase and for recovery from DNA damage. Here we report that cells lacking Slx4 have difficulties in completing DNA synthesis during recovery from replisome stalling induced by the DNA alkylating agent methyl methanesulfonate (MMS). Although DNA synthesis restarts during recovery, cells are left with unreplicated gaps in the genome despite an increase in translesion synthesis. In this light, epistasis experiments show that SLX4 interacts with genes involved in error-free bypass of DNA lesions. Slx4 associates physically, in a mutually exclusive manner, with two structure-specific endonucleases, Rad1 and Slx1, but neither of these enzymes is required for Slx4 to promote resistance to MMS. However, Rad1-dependent DNA repair by single-strand annealing (SSA) requires Slx4. Strikingly, phosphorylation of Slx4 by the Mec1 and Tel1 kinases appears to be essential for SSA but not for cell viability in the absence of Sgs1 or for cellular resistance to MMS. These results indicate that Slx4 has multiple functions in responding to DNA damage and that a subset of these are regulated by Mec1/Tel1-dependent phosphorylation.
    Original languageEnglish
    Pages (from-to)6433-45
    Number of pages13
    JournalMolecular and Cellular Biology
    Volume27
    Issue number18
    DOIs
    Publication statusPublished - Sep 2007

    Fingerprint

    Saccharomycetales
    Methyl Methanesulfonate
    DNA Repair
    Phosphorylation
    DNA
    DNA Damage
    Cell Survival
    Endonucleases
    Alkylating Agents
    Saccharomyces cerevisiae
    Phosphotransferases
    Genome
    Enzymes
    Genes

    Keywords

    • DNA Repair
    • DNA, Fungal
    • Endodeoxyribonucleases
    • Fungal Proteins
    • Intracellular Signaling Peptides and Proteins
    • Methyl Methanesulfonate
    • Models, Biological
    • Phosphorylation
    • Protein-Serine-Threonine Kinases
    • Saccharomyces cerevisiae
    • Saccharomyces cerevisiae Proteins

    Cite this

    Flott, Sonja ; Alabert, Constance ; Toh, Geraldine W ; Toth, Rachel ; Sugawara, Neal ; Campbell, David G ; Haber, James E ; Pasero, Philippe ; Rouse, John. / Phosphorylation of Slx4 by Mec1 and Tel1 regulates the single-strand annealing mode of DNA repair in budding yeast. In: Molecular and Cellular Biology. 2007 ; Vol. 27, No. 18. pp. 6433-45.
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    abstract = "Budding yeast (Saccharomyces cerevisiae) Slx4 is essential for cell viability in the absence of the Sgs1 helicase and for recovery from DNA damage. Here we report that cells lacking Slx4 have difficulties in completing DNA synthesis during recovery from replisome stalling induced by the DNA alkylating agent methyl methanesulfonate (MMS). Although DNA synthesis restarts during recovery, cells are left with unreplicated gaps in the genome despite an increase in translesion synthesis. In this light, epistasis experiments show that SLX4 interacts with genes involved in error-free bypass of DNA lesions. Slx4 associates physically, in a mutually exclusive manner, with two structure-specific endonucleases, Rad1 and Slx1, but neither of these enzymes is required for Slx4 to promote resistance to MMS. However, Rad1-dependent DNA repair by single-strand annealing (SSA) requires Slx4. Strikingly, phosphorylation of Slx4 by the Mec1 and Tel1 kinases appears to be essential for SSA but not for cell viability in the absence of Sgs1 or for cellular resistance to MMS. These results indicate that Slx4 has multiple functions in responding to DNA damage and that a subset of these are regulated by Mec1/Tel1-dependent phosphorylation.",
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    Phosphorylation of Slx4 by Mec1 and Tel1 regulates the single-strand annealing mode of DNA repair in budding yeast. / Flott, Sonja; Alabert, Constance; Toh, Geraldine W; Toth, Rachel; Sugawara, Neal; Campbell, David G; Haber, James E; Pasero, Philippe; Rouse, John.

    In: Molecular and Cellular Biology, Vol. 27, No. 18, 09.2007, p. 6433-45.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Phosphorylation of Slx4 by Mec1 and Tel1 regulates the single-strand annealing mode of DNA repair in budding yeast

    AU - Flott, Sonja

    AU - Alabert, Constance

    AU - Toh, Geraldine W

    AU - Toth, Rachel

    AU - Sugawara, Neal

    AU - Campbell, David G

    AU - Haber, James E

    AU - Pasero, Philippe

    AU - Rouse, John

    PY - 2007/9

    Y1 - 2007/9

    N2 - Budding yeast (Saccharomyces cerevisiae) Slx4 is essential for cell viability in the absence of the Sgs1 helicase and for recovery from DNA damage. Here we report that cells lacking Slx4 have difficulties in completing DNA synthesis during recovery from replisome stalling induced by the DNA alkylating agent methyl methanesulfonate (MMS). Although DNA synthesis restarts during recovery, cells are left with unreplicated gaps in the genome despite an increase in translesion synthesis. In this light, epistasis experiments show that SLX4 interacts with genes involved in error-free bypass of DNA lesions. Slx4 associates physically, in a mutually exclusive manner, with two structure-specific endonucleases, Rad1 and Slx1, but neither of these enzymes is required for Slx4 to promote resistance to MMS. However, Rad1-dependent DNA repair by single-strand annealing (SSA) requires Slx4. Strikingly, phosphorylation of Slx4 by the Mec1 and Tel1 kinases appears to be essential for SSA but not for cell viability in the absence of Sgs1 or for cellular resistance to MMS. These results indicate that Slx4 has multiple functions in responding to DNA damage and that a subset of these are regulated by Mec1/Tel1-dependent phosphorylation.

    AB - Budding yeast (Saccharomyces cerevisiae) Slx4 is essential for cell viability in the absence of the Sgs1 helicase and for recovery from DNA damage. Here we report that cells lacking Slx4 have difficulties in completing DNA synthesis during recovery from replisome stalling induced by the DNA alkylating agent methyl methanesulfonate (MMS). Although DNA synthesis restarts during recovery, cells are left with unreplicated gaps in the genome despite an increase in translesion synthesis. In this light, epistasis experiments show that SLX4 interacts with genes involved in error-free bypass of DNA lesions. Slx4 associates physically, in a mutually exclusive manner, with two structure-specific endonucleases, Rad1 and Slx1, but neither of these enzymes is required for Slx4 to promote resistance to MMS. However, Rad1-dependent DNA repair by single-strand annealing (SSA) requires Slx4. Strikingly, phosphorylation of Slx4 by the Mec1 and Tel1 kinases appears to be essential for SSA but not for cell viability in the absence of Sgs1 or for cellular resistance to MMS. These results indicate that Slx4 has multiple functions in responding to DNA damage and that a subset of these are regulated by Mec1/Tel1-dependent phosphorylation.

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    KW - Endodeoxyribonucleases

    KW - Fungal Proteins

    KW - Intracellular Signaling Peptides and Proteins

    KW - Methyl Methanesulfonate

    KW - Models, Biological

    KW - Phosphorylation

    KW - Protein-Serine-Threonine Kinases

    KW - Saccharomyces cerevisiae

    KW - Saccharomyces cerevisiae Proteins

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    EP - 6445

    JO - Molecular and Cellular Biology

    JF - Molecular and Cellular Biology

    SN - 0270-7306

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    ER -