TPR proteins required for anaphase progression mediate ubiquitination of mitotic B-type cyclins in yeast.
The abundance of B-type cyclin-CDK complexes is determined by regulated synthesis and degradation of cyclin subunits. Cyclin proteolysis is required for the final exit from mitosis and for the initiation of a new cell cycle. In extracts from frog or clam eggs, degradation is accompanied by ubiquitination of cyclin. Three genes, CDC16, CDC23, and CSE1 have recently been shown to be required specifically for cyclin B proteolysis in yeast. To test whether these genes are required for cyclin ubiquitination, we prepared extracts from G1-arrested yeast cells capable of conjugating ubiquitin to the B-type cyclin Clb2. The ubiquitination activity was cell cycle regulated, required Clb2's destruction box, and was low if not absent in cdc16, cdc23, cdc27, and cse1 mutants. Furthermore all these mutants were also defective in ubiquitination of another mitotic B-type cyclin, Clb3. The Cdc16, Cdc23, and Cdc27 proteins all contain several copies of the tetratricopeptide repeat and are subunits of a complex that is required for the onset of anaphase. The finding that gene products that are required for ubiquitination of Clb2 and Clb3 are also required for cyclin proteolysis in vivo provides the best evidence so far that cyclin B is degraded via the ubiquitin pathway in living cells. Xenopus homologues of Cdc16 and Cdc27 have meanwhile been shown to be associated with a 20S particle that appears to function as a cell cycle-regulated ubiquitin-protein ligase.
Zachariae
W
W
Research Institute of Molecular Pathology, Vienna, Austria.
Nasmyth
K
K
D000705:Anaphase / Q000502:physiology*; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D016213:Cyclins / Q000502:physiology*; D005434:Flow Cytometry; D016193:G1 Phase / Q000235:genetics; D010957:Plasmids; D014452:Ubiquitins / Q000378:metabolism*; D015003:Yeasts / Q000502:physiology*
D000911:Antibodies, Monoclonal; D016213:Cyclins; D014452:Ubiquitins