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Vol. 14, No. 18, pp. 2344-2357, September 15, 2000

RESEARCH PAPER
A nonproteolytic function of the proteasome is required for the dissociation of Cdc2 and cyclin B at the end of M phase

Atsuya Nishiyama,1 Kazunori Tachibana,1 Yuko Igarashi,2 Hideyo Yasuda,2 Nobuyuki Tanahashi,3 Keiji Tanaka,3,4 Keita Ohsumi,1 and Takeo Kishimoto1,4,5

1 Laboratory of Cell and Developmental Biology, Graduate School of Biosciences, Tokyo Institute of Technology, Midori-ku, Yokohama 226-8501, Japan; 2 School of Life Science, Tokyo University of Pharmacy, Hachiooji, Tokyo 192-0355, Japan; 3 Tokyo Metropolitan Institute of Medical Science, Bunkyo-ku, Tokyo 113-8613, Japan; 4 CREST Research Project, Japan Science and Technology Corporation, Japan

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.

[Key Words: cyclin B-Cdc2; proteasome; cell cycle; protein unfolding; Xenopus egg extracts]

5 Corresponding author.

GENES & DEVELOPMENT 14:2344-2357 © 2000 by Cold Spring Harbor Laboratory Press  ISSN 0890-9369/00 $5.00
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