Ca(2+)-promoted cyclin B1 degradation in mouse oocytes requires the establishment of a metaphase arrest.
CDK1-cyclin B1 is a universal cell cycle kinase required for mitotic/meiotic cell cycle entry and its activity needs to decline for mitotic/meiotic exit. During their maturation, mouse oocytes proceed through meiosis I and arrest at second meiotic metaphase with high CDK1-cyclin B1 activity. Meiotic arrest is achieved by the action of a cytostatic factor (CSF), which reduces cyclin B1 degradation. Meiotic arrest is broken by a Ca2+ signal from the sperm that accelerates it. Here we visualised degradation of cyclin B1::GFP in oocytes and found that its degradation rate was the same for both meiotic divisions. Ca2+ was the necessary and sufficient trigger for cyclin B1 destruction during meiosis II; but it played no role during meiosis I and furthermore could not accelerate cyclin B1 destruction during this time. The ability of Ca2+ to trigger cyclin B1 destruction developed in oocytes following a restabilisation of cyclin B1 levels at about 12 h of culture. This was independent of actual first polar body extrusion. Thus, in metaphase I arrested oocytes, Ca2+ would induce cyclin B1 destruction and the first polar body would be extruded. In contrast to some reports in lower species, we found no evidence that oocyte activation was associated with an increase in 26S proteasome activity. We therefore conclude that Ca2+ mediates cyclin B1 degradation by increasing the activity of an E3 ubiquitin ligase. However, this stimulation occurs only in the presence of the ubiquitin ligase inhibitor CSF. We propose a model in which Ca2+ directly stimulates destruction of CSF during mammalian fertilisation.
Hyslop
Louise A
LA
Cell and Developmental Physiology Research Group, School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK.
Nixon
Victoria L
VL
Levasseur
Mark
M
Chapman
Faye
F
Chiba
Kazuyoshi
K
McDougall
Alex
A
Venables
Julian P
JP
Elliott
David J
DJ
Jones
Keith T
KT
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002842:Chromatids / Q000378:metabolism; D020090:Chromosome Segregation / Q000502:physiology; D019926:Cyclin B / Q000378:metabolism*; D056744:Cyclin B1; D005260:Female; D008540:Meiosis / Q000502:physiology; D008677:Metaphase / Q000502:physiology; D051379:Mice; D009865:Oocytes / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex
C534700:Ccnb1 protein, mouse; D019926:Cyclin B; D056744:Cyclin B1; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D002118:Calcium