Construction by gene targeting in human cells of a "conditional' CDC2 mutant that rereplicates its DNA.
We describe a novel gene targeting strategy for the genetic analysis of essential genes in mammalian cells and its use to study the role of the cell cycle control gene CDC2 in human cells. A cell line (HT2-19) was generated in which endogenous CDC2 gene expression and cell viability depend on the presence of an inducer in the growth medium. In the absence of inducer, HT2-19 cells undergo extensive DNA rereplication and apoptosis. Rereplication is indicative of a role for human CDC2 in a control mechanism, previously undetected in mammalian cells, that prevents premature entry into S-phase.
Itzhaki
J E
JE
MRC Clinical Sciences Centre, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.
Gilbert
C S
CS
Porter
A C
AC
D017209:Apoptosis; D001426:Bacterial Proteins / Q000096:biosynthesis; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000235:genetics* / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D017931:DNA Primers; D004261:DNA Replication; D029968:Escherichia coli Proteins; D005816:Genetic Complementation Test; D005821:Genetic Techniques; D006801:Humans; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D056968:Lac Repressors; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012097:Repressor Proteins / Q000096:biosynthesis; D016196:S Phase; D014162:Transfection
D001426:Bacterial Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; D056968:Lac Repressors; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D007544:Isopropyl Thiogalactoside; D016203:CDC2 Protein Kinase