DNA-Binding Proteins; Saccharomyces cerevisiae Proteins; Schizosaccharomyces pombe Proteins; Eme1 protein, S pombe; Eme1 protein, mouse; Endodeoxyribonucleases; Endonucleases; MUS81 protein, S cerevisiae; MUS81 protein, human; Mus81 protein, mouse; Amino Acid Sequence; Animals; Chromosomal Instability; DNA Damage; DNA Repair/physiology; DNA-Binding Proteins/metabolism; Endodeoxyribonucleases/genetics; Endodeoxyribonucleases/metabolism; Endonucleases/genetics; Endonucleases/metabolism; Humans; Mice; Molecular Sequence Data; Schizosaccharomyces pombe Proteins/genetics; Sister Chromatid Exchange; Stem Cells; Genomic Instability; DNA repair; Homologous recombination; Inter-strand DNA cross-links; Mus81-Eme1; Neuroscience (all); Molecular Biology; Biochemistry, Genetics and Molecular Biology (all); Immunology and Microbiology (all)
Abstract :
[en] Yeast and human Eme1 protein, in complex with Mus81, constitute an endonuclease that cleaves branched DNA structures, especially those arising during stalled DNA replication. We identified mouse Eme1, and show that it interacts with Mus81 to form a complex that preferentially cleaves 3'-flap structures and replication forks rather than Holliday junctions in vitro. We demonstrate that Eme1-/- embryonic stem (ES) cells are hypersensitive to the DNA cross-linking agents mitomycin C and cisplatin, but only mildly sensitive to ionizing radiation, UV radiation and hydroxyurea treatment. Mammalian Eme1 is not required for the resolution of DNA intermediates that arise during homologous recombination processes such as gene targeting, gene conversion and sister chromatid exchange (SCE). Unlike Blm-deficient ES cells, increased SCE was seen only following induced DNA damage in Eme1-deficient cells. Most importantly, Eme1 deficiency led to spontaneous genomic instability. These results reveal that mammalian Eme1 plays a key role in DNA repair and the maintenance of genome integrity.
Disciplines :
Genetics & genetic processes
Author, co-author :
Abraham, Jacinth; Advanced Medical Discovery Institute, Ontario Cancer Institute, 620 University Avenue, Suite 706, Toronto, Ontario M5G 2C1, Canada
Lemmers, Bénédicte; Advanced Medical Discovery Institute, Ontario Cancer Institute, Toronto, Ont. M5G 2C1, Canada ; Department of Medical Biophysics, University of Toronto, Toronto, Ont., Canada
Hande, M Prakash; Faculty of Medicine, National University of Singapore, Singapore 117597, Singapore
Moynahan, Mary Ellen; Department of Medicine, Mem. Sloan-Kettering Cancer Center, New York, NY 10021, United States
Chahwan, Charly; Advanced Medical Discovery Institute, Ontario Cancer Institute, Toronto, Ont. M5G 2C1, Canada
Ciccia, Alberto; Cancer Research UK, London Research Institute, Clare Hall Laboratories, South Mimms, Herts EN6 3LD, United Kingdom
Essers, Jeroen; Dept. of Cell Biology and Genetics, Erasmus MC, 3000 DR Rotterdam, Netherlands
Hanada, Katsuhiro; Dept. of Cell Biology and Genetics, Erasmus MC, 3000 DR Rotterdam, Netherlands
CHAHWAN, Richard ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Health, Medicine and Life Sciences (DHML) ; Advanced Medical Discovery Institute, Ontario Cancer Institute, Toronto, Ont. M5G 2C1, Canada
Khaw, Aik Kia; Faculty of Medicine, National University of Singapore, Singapore 117597, Singapore
McPherson, Peter; Advanced Medical Discovery Institute, Ontario Cancer Institute, Toronto, Ont. M5G 2C1, Canada
Shehabeldin, Amro; Advanced Medical Discovery Institute, Ontario Cancer Institute, Toronto, Ont. M5G 2C1, Canada ; Department of Medical Biophysics, University of Toronto, Toronto, Ont., Canada
Laister, Rob; Department of Medical Biophysics, University of Toronto, Toronto, Ont., Canada
Arrowsmith, Cheryl; Department of Medical Biophysics, University of Toronto, Toronto, Ont., Canada
Kanaar, Roland; Dept. of Cell Biology and Genetics, Erasmus MC, 3000 DR Rotterdam, Netherlands
West, Stephen C; Cancer Research UK, London Research Institute, Clare Hall Laboratories, South Mimms, Herts EN6 3LD, United Kingdom
Jasin, Maria; Department of Medicine, Mem. Sloan-Kettering Cancer Center, New York, NY 10021, United States
Hakem, Razqallah; Advanced Medical Discovery Institute, Ontario Cancer Institute, Toronto, Ont. M5G 2C1, Canada ; Department of Medical Biophysics, University of Toronto, Toronto, Ont., Canada
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