Identification of MMS22 as a regulator of DNA repair

  • Eris Duro

    Student thesis: Doctoral ThesisDoctor of Philosophy

    Abstract

    Obstacles such as DNA damage can block the progression of DNA replication forks. This is a major source of genome instability that can lead to cell transformation or death. The budding yeast MMS1 and MMS22 genes were identified in a screen for mutants that were hypersensitive to DNA alkylation that blocks replisome progression. I set out to investigate the cellular roles of these genes and found that cells lacking MMS1 or MMS22 are hypersensitive to a wide variety of genotoxins that stall or block replication forks, and are severely defective in their ability to recover from DNA alkylation damage. Homologous recombination (HR) is an important mechanism for the rescue of stalled or blocked replication forks and for the repair of double-strand breaks (DSBs). Strikingly, MMS1 and MMS22 are required for HR induced by replication stress but not by DSBs, and the underlying mechanisms were explored.I next identified the uncharacterized protein C6ORF167 (MMS22L) as a putative human Mms22 orthologue. MMS22L interacts with NF?BIL2/TONSL, the histone chaperone ASF1 and subunits of the MCM replicative helicase. MMS22L colocalizes with TONSL at perturbed replication forks and at sites of DNA damage. MMS22L and TONSL are important for the repair of collapsed replication forks as depletion of MMS22L or TONSL from human cells causes DNA damage during S–phase and hypersensitivity to agents that cause fork collapse. These defects are consistent with the observations that MMS22L and TONSL are required for the efficient loading of the RAD51 recombinase onto resected DNA ends and for efficient HR. These data indicate that MMS22L and TONSL are novel regulators of genome stability that enable efficient HR.
    Date of Award2010
    Original languageEnglish
    Awarding Institution
    • University of Dundee
    SponsorsMedical Research Council & Dorothy Hodgkin Postgraduate Award
    SupervisorJohn Rouse (Supervisor)

    Keywords

    • DNA damage
    • Genome stability
    • Homologous recombination
    • MMS22
    • MMS22L
    • NFkBIL2

    Cite this

    Identification of MMS22 as a regulator of DNA repair
    Duro, E. (Author). 2010

    Student thesis: Doctoral ThesisDoctor of Philosophy