Analysis of the role of phosphorylation in fission yeast Cdc13p/cyclinB function.
The Cdk1p-cyclin B complex drives entry into mitosis in all eukaryotes. Cdc13p is the single essential cyclin in Schizosaccharomyces pombe and a member of the cyclin B family. Cdc13p abundance rises during G(2)-phase and falls as cells progress through mitosis and G(1). Cdc13p degradation, mediated by the anaphase-promoting complex, is an important mechanism of Cdk1p inhibition and mitotic exit. Cdk1p-cyclin B1 complexes shuttle between the nucleus and cytoplasm, and preventing nuclear accumulation of Cdk1p-cyclin B1 in mammalian cells appears to be one mechanism of preventing entry into mitosis during a DNA damage-induced checkpoint delay. In vertebrates, phosphorylation plays a key role in regulating the intracellular distribution of cyclins. Previous mass spectrometric analysis identified sites of Cdc13p phosphorylation. Here, we have confirmed that these sites are the sole in vivo Cdc13p phosphorylation sites and have studied the role that phosphorylation plays in Cdc13p localization and function. Our data indicate that Cdc13p accumulates in the nucleolus in response to G(2) checkpoint delays, rather than in the cytoplasm, and that phosphorylation plays no role in Cdc13p localization or function.
Ren
Liping
L
Howard Hughes Medical Institute and Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Feoktistova
Anna
A
McDonald
W Hayes
WH
Haese
Greg Den
GD
Morrell
Jennifer L
JL
Gould
Kathleen L
KL
D000483:Alleles; D001665:Binding Sites; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism; D019926:Cyclin B / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004249:DNA Damage; D004591:Electrophoresis, Polyacrylamide Gel; D016193:G1 Phase; D016195:G2 Phase; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008938:Mitosis; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010766:Phosphorylation; D012568:Schizosaccharomyces / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D015398:Signal Transduction
D019926:Cyclin B; D049452:Green Fluorescent Proteins; D012694:Serine