A deletion within the murine Werner syndrome helicase induces sensitivity to inhibitors of topoisomerase and loss of cellular proliferative capacity.
Werner syndrome (WS) is an autosomal recessive disorder characterized by genomic instability and the premature onset of a number of age-related diseases. The gene responsible for WS encodes a member of the RecQ-like subfamily of DNA helicases. Here we show that its murine homologue maps to murine chromosome 8 in a region syntenic with the human WRN gene. We have deleted a segment of this gene and created Wrn-deficient embryonic stem (ES) cells and WS mice. While displaying reduced embryonic survival, live-born WS mice otherwise appear normal during their first year of life. Nonetheless, although several DNA repair systems are apparently intact in homozygous WS ES cells, such cells display a higher mutation rate and are significantly more sensitive to topoisomerase inhibitors (especially camptothecin) than are wild-type ES cells. Furthermore, mouse embryo fibroblasts derived from homozygous WS embryos show premature loss of proliferative capacity. At the molecular level, wild-type, but not mutant, WS protein copurifies through a series of centrifugation and chromatography steps with a multiprotein DNA replication complex.
Lebel
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M
Department of Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.
Leder
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P
D000818:Animals; D001483:Base Sequence; D002166:Camptothecin / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002678:Chimera; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D004265:DNA Helicases / Q000235:genetics*; D017931:DNA Primers; D005260:Female; D005313:Fetal Death; D006720:Homozygote; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D009115:Muridae; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011995:Recombination, Genetic; D017384:Sequence Deletion; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D014162:Transfection; D014898:Werner Syndrome / Q000201:enzymology / Q000235:genetics*
D017931:DNA Primers; D004265:DNA Helicases; D002166:Camptothecin