A nonproteolytic function of the proteasome is required for the dissociation of Cdc2 and cyclin B at the end of M phase.
Inactivation of cyclin B-Cdc2 kinase at the exit from M phase depends on the specific proteolysis of the cyclin B subunit, whereas the Cdc2 subunit remains present at nearly constant levels throughout the cell cycle. It is unknown how Cdc2 escapes degradation when cyclin B is destroyed. In Xenopus egg extracts that reproduce the exit from M phase in vitro, we have found that dissociation of the cyclin B-Cdc2 complex occurred under conditions where cyclin B was tethered to the 26S proteasome but not yet degraded. The dephosphorylation of Thr 161 on Cdc2 was unlikely to be necessary for the dissociation of the two subunits. However, the dissociation was dependent on the presence of a functional destruction box in cyclin B. Cyclin B ubiquitination was also, by itself, not sufficient for separation of Cdc2 and cyclin B. The 26S proteasome, but not the 20S proteasome, was capable of dissociating the two subunits. These results indicate that the cyclin B and Cdc2 subunits are separated by the proteasome through a mechanism that precedes proteolysis of cyclin B and is independent of proteolysis. As a result, cyclin B levels decrease on exit from M phase but Cdc2 levels remain constant.
Nishiyama
A
A
Laboratory of Cell and Developmental Biology, Graduate School of Biosciences, Tokyo Institute of Technology, Midori-ku, Yokohama 226-8501, Japan.
Tachibana
K
K
Igarashi
Y
Y
Yasuda
H
H
Tanahashi
N
N
Tanaka
K
K
Ohsumi
K
K
Kishimoto
T
T
D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D019926:Cyclin B / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D066298:In Vitro Techniques; D008938:Mitosis; D009097:Multienzyme Complexes / Q000378:metabolism*; D010063:Ovum / Q000254:growth & development / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D011233:Precipitin Tests; D006656:Protamine Kinase / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D014452:Ubiquitins / Q000378:metabolism; D014981:Xenopus
D019926:Cyclin B; D009097:Multienzyme Complexes; D014452:Ubiquitins; D006656:Protamine Kinase; D016203:CDC2 Protein Kinase; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease