Differential requirements for DNA replication in the activation of mitotic checkpoints in Saccharomyces cerevisiae.
Checkpoints prevent inaccurate chromosome segregation by inhibiting cell division when errors in mitotic processes are encountered. We used a temperature-sensitive mutation, dbf4, to examine the requirement for DNA replication in establishing mitotic checkpoint arrest. We used gamma-irradiation to induce DNA damage and hydroxyurea to limit deoxyribonucleotides in cells deprived of DBF4 function to investigate the requirement for DNA replication in DNA-responsive checkpoints. In the absence of DNA replication, mitosis was not inhibited by these treatments, which normally activate the DNA damage and DNA replication checkpoints. Our results support a model that indicates that the assembly of replication structures is critical for cells to respond to defects in DNA metabolism. We show that activating the spindle checkpoint with nocodazole does not require prior progression through S phase but does require a stable kinetochore.
Tavormina
P A
PA
Department of Biology, University of Virginia, Charlottesville 22903, USA.
Wang
Y
Y
Burke
D J
DJ
D000970:Antineoplastic Agents / Q000494:pharmacology; D018797:Cell Cycle Proteins; D002455:Cell Division; D002842:Chromatids / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004261:DNA Replication; D004271:DNA, Fungal / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism; D008938:Mitosis / Q000502:physiology*; D015739:Nocodazole / Q000494:pharmacology; D009687:Nuclear Proteins / Q000378:metabolism; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D008941:Spindle Apparatus / Q000378:metabolism
D000970:Antineoplastic Agents; C081354:CTF13 protein, S cerevisiae; D018797:Cell Cycle Proteins; D004271:DNA, Fungal; C088096:Dbf4 protein, S cerevisiae; D005656:Fungal Proteins; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; D015739:Nocodazole