Efficient conditional mutation of the vertebrate CENP-C gene.
We have used gene targeting in the DT40 cell line to create a cell line which expresses a fusion between CENP-C and a mouse steroid receptor and which behaves as a conditional loss of function mutant of CENP-C. Under restrictive conditions these cells arrest at the metaphase/anaphase junction and after a delay of approximately 2.5 h die by apoptosis. These results indicate that CENP-C is either necessary for anaphase chromosome movement or for mediating a signal which triggers centromere function during anaphase. Our approach is simple and applicable to a wide range of proteins with general cell autonomous functions in vertebrates.
Fukagawa
T
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Cancer Research Campaign Chromosome Molecular Biology Group, Biochemistry Department, Oxford University, South Parks Road, Oxford OX1 3QU, UK. tfukagaw@bioch.ox.ac.uk
Brown
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WR
D000595:Amino Acid Sequence; D000705:Anaphase / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002503:Centromere / Q000737:chemistry / Q000502:physiology; D002645:Chickens / Q000235:genetics; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics* / Q000502:physiology; D018390:Gene Targeting; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011955:Receptors, Drug / Q000187:drug effects / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology; D014162:Transfection; D014714:Vertebrates / Q000235:genetics*
D002868:Chromosomal Proteins, Non-Histone; D011955:Receptors, Drug; D011993:Recombinant Fusion Proteins; C060905:centromere protein C; D013629:Tamoxifen; C016601:afimoxifene