Activation of p34cdc2 kinase by cyclin A.
Functional clam cyclin A and B proteins have been produced using a baculovirus expression system. Both cyclin A and B can induce meiosis I and meiosis II in Xenopus in the absence of protein synthesis. Half-maximal induction occurs at 50 nM for cyclin A and 250 nM for cyclin B. Addition of 25 nM cyclin A to activated Xenopus egg extracts arrested in the cell cycle by treatment with RNase or emetine activates cdc2 kinase to the normal metaphase level and stimulates one oscillatory cell cycle. High levels of cyclin A cause marked hyperactivation of cdc2 kinase and a stable arrest at the metaphase point in the cell cycle. Kinetic studies demonstrate the concentration of cyclin A added does not affect the 10 min lag period required for kinase activation or the timing of maximal activity, but does control the rate of deactivation of cdc2 kinase during exit from mitosis. In addition, exogenous clam cyclin A inhibits the degradation of both A- and B-type endogenous Xenopus cyclins. These results define a system for investigating the biochemistry and regulation of cdc2 kinase activation by cyclin A.
Roy
L M
LM
Howard Hughes Medical Institute, University of Colorado School of Medicine, Denver 80262.
Swenson
K I
KI
Walker
D H
DH
Gabrielli
B G
BG
Li
R S
RS
Piwnica-Worms
H
H
Maller
J L
JL
D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002460:Cell Line; D016213:Cyclins / Q000378:metabolism / Q000494:pharmacology*; D003513:Cycloheximide / Q000494:pharmacology; D004789:Enzyme Activation; D016200:Maturation-Promoting Factor / Q000378:metabolism; D008540:Meiosis / Q000187:drug effects; D008938:Mitosis / Q000187:drug effects; D009865:Oocytes; D014176:Protein Biosynthesis; D014982:Xenopus laevis
D016213:Cyclins; D003513:Cycloheximide; D016203:CDC2 Protein Kinase; D016200:Maturation-Promoting Factor