Electronic versions

DOI

  • E. Hartsuiker
    University of Sussex
  • E. Vaessen
    Fribourg University
  • A.M. Carr
    University of Sussex
  • J. Kohli
    Bern University

To study the role of Rad50 in the DNA damage response, we cloned and deleted the Schizosaccharomyces pombe RAD50 homologue. The deletion is sensitive to a range of DNA-damaging agents and shows dynamic epistatic interactions with other recombination-repair genes. We show that Rad50 is necessary for recombinational repair of the DNA lesion at the mating-type locus and that rad50Delta shows slow DNA replication. We also find that Rad50 is not required for slowing down S phase in response to hydroxy urea or methyl methanesulfonate (MMS) treatment. Interestingly, in rad50Delta cells, the recombination frequency between two homologous chromosomes is increased at the expense of sister chromatid recombination. We propose that Rad50, an SMC-like protein, promotes the use of the sister chromatid as the template for homologous recombinational repair. In support of this, we found that Rad50 functions in the same pathway for the repair of MMS-induced damage as Rad21, the homologue of the Saccharomyces cerevisiae Scc1 cohesin protein. We speculate that Rad50 interacts with the cohesin complex during S phase to assist repair and possibly re-initiation of replication after replication fork collapse.

Keywords

  • Amino Acid Sequence, Animals, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Cell Cycle Proteins, Cell Separation, Chromosomal Proteins, Non-Histone, Chromosome Deletion, Cloning, Molecular, Crosses, Genetic, DNA Damage, DNA Repair, DNA Replication, DNA-Binding Proteins, Dose-Response Relationship, Drug, Dose-Response Relationship, Radiation, Epistasis, Genetic, Flow Cytometry, Fungal Proteins, Gamma Rays, Gene Deletion, Genotype, Humans, Hydroxyurea, Methyl Methanesulfonate, Mice, Models, Genetic, Molecular Sequence Data, Nuclear Proteins, Phosphoproteins, Recombination, Genetic, S Phase, Saccharomyces cerevisiae Proteins, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Sequence Homology, Amino Acid, Sister Chromatid Exchange, Telomere, Time Factors, Journal Article, Research Support, Non-U.S. Gov't
Original languageEnglish
Pages (from-to)6660-71
Number of pages12
JournalEMBO Journal
Volume20
Issue number23
DOIs
Publication statusPublished - 3 Dec 2001
Externally publishedYes
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