Spatial regulation of APCCdh1-induced cyclin B1 degradation maintains G2 arrest in mouse oocytes.
Within the mammalian ovary, oocytes remain arrested at G2 for several years. Then a peri-ovulatory hormonal cue triggers meiotic resumption by releasing an inhibitory phosphorylation on the kinase Cdk1. G2 arrest, however, also requires control in the concentrations of the Cdk1-binding partner cyclin B1, a process achieved by anaphase-promoting complex (APC(Cdh1)) activity, which ubiquitylates and so targets cyclin B1 for degradation. Thus, APC(Cdh1) activity prevents precocious meiotic entry by promoting cyclin B1 degradation. However, it remains unresolved how cyclin B1 levels are suppressed sufficiently to maintain arrest but not so low that they make oocytes hormonally insensitive. Here, we examined spatial control of this process by determining the intracellular location of the proteins involved and using nuclear-targeted cyclin B1. We found that raising nuclear cyclin B1 concentrations, an event normally observed in the minutes before nuclear envelope breakdown, was a very effective method of inducing the G2/M transition. Oocytes expressed only the alpha-isoform of Cdh1, which was predominantly nuclear, as were Cdc27 and Psmd11, core components of the APC and the 26S proteasome, respectively. Furthermore, APC(Cdh1) activity appeared higher in the nucleus, as nuclear-targeted cyclin B1 was degraded at twice the rate of wild-type cyclin B1. We propose a simple spatial model of G2 arrest in which nuclear APC(Cdh1)-proteasomal activity guards against any cyclin B1 accumulation mediated by nuclear import.
School of Biomedical Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
Janet E
JE
Holt
Jessica
J
Weaver
Keith T
KT
Jones
D017398:Alternative Splicing; D064173:Anaphase-Promoting Complex-Cyclosome; D000818:Animals; D000938:Antigen-Presenting Cells / Q000166:cytology / Q000502:physiology; D016203:CDC2 Protein Kinase / Q000235:genetics / Q000378:metabolism*; D064200:Cdh1 Proteins; D018797:Cell Cycle Proteins / Q000235:genetics; D002455:Cell Division / Q000502:physiology; D002467:Cell Nucleus / Q000502:physiology; D056744:Cyclin B1 / Q000378:metabolism*; D005260:Female; D016195:G2 Phase / Q000502:physiology; D005786:Gene Expression Regulation; D008540:Meiosis; D051379:Mice; D008938:Mitosis / Q000502:physiology; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D010053:Ovary / Q000166:cytology / Q000502:physiology; D014158:Transcription, Genetic; D043743:Ubiquitin-Protein Ligase Complexes / Q000502:physiology*
D064200:Cdh1 Proteins; D018797:Cell Cycle Proteins; D056744:Cyclin B1; C577585:Fzr1 protein, mouse; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome; D016203:CDC2 Protein Kinase