Purification of the cdc8 protein of Saccharomyces cerevisiae by complementation in an aphidicolin-sensitive in vitro DNA replication system.
DNA synthesis in vitro in Brij-treated Saccharomyces cerevisiae requires the product of the CDC8 gene (Hereford, L. M. & Hartwell, L. H. (1971) Nature (London) New Biol. 234, 171-172). Extracts of wild-type A364a yeast restore DNA synthesis in Brij-treated cdc8, a mutant containing a thermolabile cdc8 gene product. This constitutes a complementation assay by which the cdc8 gene product can be monitored during purification. A heat-stable protein responsible for this complementation has been partially purified from both wild-type A364a cells and from a cdc8 temperature-sensitive mutant. The complementation activity from the mutant is thermolabile when compared to the wild-type activity, indicating that CDC8 is the structural gene for the protein.
D016590:Aphidicolin; D002453:Cell Cycle; D002463:Cell Membrane Permeability; D004261:DNA Replication / Q000187:drug effects; D004271:DNA, Fungal / Q000096:biosynthesis; D004224:Diterpenes / Q000494:pharmacology; D005656:Fungal Proteins / Q000302:isolation & purification*; D005816:Genetic Complementation Test; D006358:Hot Temperature; D019384:Nucleic Acid Synthesis Inhibitors; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D004271:DNA, Fungal; D004224:Diterpenes; D005656:Fungal Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D016590:Aphidicolin