A novel yeast screen for mitotic arrest mutants identifies DOC1, a new gene involved in cyclin proteolysis.
B-type cyclins are rapidly degraded at the transition between metaphase and anaphase and their ubiquitin-mediated proteolysis is required for cells to exit mitosis. We used a novel enrichment to isolate new budding mutants that arrest the cell cycle in mitosis. Most of these mutants lie in the CDC16, CDC23, and CDC27 genes, which have already been shown to play a role in cyclin proteolysis and encode components of a 20S complex (called the cyclosome or anaphase promoting complex) that ubiquitinates mitotic cyclins. We show that mutations in CDC26 and a novel gene, DOC1, also prevent mitotic cyclin proteolysis. Mutants in either gene arrest as large budded cells with high levels of the major mitotic cyclin (Clb2) protein at 37 degrees C and cannot degrade Clb2 in G1-arrested cells. Cdc26 associates in vivo with Doc1, Cdc16, Cdc23, and Cdc27. In addition, the majority of Doc1 cosediments at 20S with Cdc27 in a sucrose gradient, indicating that Cdc26 and Doc1 are components of the anaphase promoting complex.
Hwang
L H
LH
Department of Physiology, University of California, San Francisco 94143-0444, USA.
Murray
A W
AW
D000595:Amino Acid Sequence; D064197:Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome; D002453:Cell Cycle / Q000502:physiology; D018797:Cell Cycle Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D016213:Cyclins / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000235:genetics; D005656:Fungal Proteins / Q000378:metabolism; D017491:Genes, APC; D005800:Genes, Fungal; D019940:Genes, bcl-1 / Q000235:genetics*; D006868:Hydrolysis; D008938:Mitosis / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics; D016296:Mutagenesis; D009154:Mutation; D046988:Proteasome Endopeptidase Complex; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid
D064197:Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome; D018797:Cell Cycle Proteins; D016213:Cyclins; C577456:DOC1 protein, S cerevisiae; D005656:Fungal Proteins; D009097:Multienzyme Complexes; D029701:Saccharomyces cerevisiae Proteins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex