Differential mitotic degradation of the CDC25B phosphatase variants.
CDC25 phosphatases control cell-cycle progression by dephosphorylating and activating cyclin-dependent kinases. CDC25B, one of the three members of this family in human cells, is thought to regulate initial mitotic events. CDC25B is an unstable protein whose proteasomal degradation is proposed to be controlled by beta-TrCP. Here, we have investigated the regulation of CDC25B during mitosis, using time-lapse video microscopy. We found that CDC25B expression is high during early mitosis, and that its degradation occurs after the metaphase-anaphase transition and cyclin B1 destruction. We also show that CDC25B degradation after metaphase is dependent on the integrity of the KEN-box and RRKSE motifs that are located within the alternatively spliced B domain, and that the CDC25B2 splice variant, that lacks this domain, is stable during mitosis. Furthermore, we show that the N-terminal region of CDC25B, encompassing the B domain, undergoes major conformational changes during mitosis that can be monitored by intramolecular fluorescence resonance energy transfer variation of specific CDC25B biosensors. This study demonstrates that CDC25B splice variants have differential mitotic stabilities, a feature that is likely to have major consequences on the local control of cyclin-dependent kinase-cyclin activities during mitotic progression.
Kieffer
I
I
LBCMCP-CNRS UMR5088-IFR109 Institut d'Exploration Fonctionnelle des Génomes, Université Paul Sabatier, 118 route de Narbonne, Toulouse, France.
Lorenzo
C
C
Dozier
C
C
Schmitt
E
E
Ducommun
B
B
D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism; D019926:Cyclin B / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D006367:HeLa Cells; D006801:Humans; D008677:Metaphase; D051379:Mice; D008938:Mitosis; D017434:Protein Structure, Tertiary; D044844:beta-Transducin Repeat-Containing Proteins / Q000378:metabolism; D020687:cdc25 Phosphatases / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D019926:Cyclin B; D044844:beta-Transducin Repeat-Containing Proteins; D016203:CDC2 Protein Kinase; C517008:CDC25B protein, human; D020687:cdc25 Phosphatases