PD 98059 prevents establishment of the spindle assembly checkpoint and inhibits the G2-M transition in meiotic but not mitotic cell cycles in Xenopus.
Most chemotherapeutic agents block DNA replication, damage DNA, or interfere with chromosome segregation. The existence of checkpoints, which monitor these events, indicates that mechanisms exist to avoid death when essential cellular events are inhibited. A molecular understanding of cellular checkpoints should therefore provide opportunities for the development of inhibitors of checkpoint controls which may increase the potency of chemotherapeutic drugs by inducing catastrophic cell cycle progression. The molecular dissection of cell cycle arrest points is facilitated in the Xenopus egg/oocyte system, in which cell-free systems retain both S/M and spindle assembly checkpoints. Members of the MAP kinase family have been shown to play a role in the induction of G2 to M transition during oocyte maturation and have been implicated in the maintenance of either cytostatic factor- or spindle assembly checkpoint-induced M-phase arrest. Here, we have examined the effects of the inhibitor of MAP kinase kinase activation, PD 98059, on cell cycle progression in Xenopus oocytes and in cell-free extracts. This inhibitor is highly specific for the kinase which activates the classical p42/p44 MAP kinase, having no effect on upstream activators of stress-activated protein kinases. We have found that PD 98059 inhibits oocyte maturation, consistent with a role for p42 MAP kinase as a rate-limiting component in the induction of meiosis, but had no effect on the timing of G2-M transition in cell-free extracts indicating that, unlike meiosis, p42 MAP kinase activation is not limiting for normal mitotic M phase entry. However, we found that cytostatic factor-induced metaphase arrest, as well as the spindle assembly checkpoint, were both abolished in the presence of the drug. These results demonstrate that p42 MAP kinase, and not some other member of the MAP kinase family, is responsible for both CSF- and checkpoint-induced metaphase arrest and suggest that PD 98059 and similar agents may have considerable therapeutic potential for the potentiation of chemotherapeutic regimes.
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Department of Biochemistry, The University, Dundee, United Kingdom.
Smythe
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D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002474:Cell-Free System / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D005419:Flavonoids / Q000494:pharmacology*; D016195:G2 Phase / Q000187:drug effects*; D008540:Meiosis / Q000187:drug effects; D020929:Mitogen-Activated Protein Kinase Kinases; D008938:Mitosis / Q000187:drug effects*; D009865:Oocytes / Q000187:drug effects / Q000201:enzymology / Q000254:growth & development; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000187:drug effects; D017231:Proto-Oncogene Proteins c-mos / Q000494:pharmacology; D008941:Spindle Apparatus / Q000187:drug effects* / Q000378:metabolism; D014982:Xenopus laevis / Q000378:metabolism*
D004791:Enzyme Inhibitors; D005419:Flavonoids; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D017231:Proto-Oncogene Proteins c-mos; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D020929:Mitogen-Activated Protein Kinase Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one