Combinatorial control of cyclin B1 nuclear trafficking through phosphorylation at multiple sites.
Entry into mitosis is regulated by the Cdc2 kinase complexed to B-type cyclins. We and others recently reported that cyclin B1/Cdc2 complexes, which appear to be constitutively cytoplasmic during interphase, actually shuttle continually into and out of the nucleus, with the rate of nuclear export exceeding the import rate (). At the time of entry into mitosis, the import rate is increased, whereas the export rate is decreased, leading to rapid nuclear accumulation of Cdc2/cyclin B1. Although it has recently been reported that phosphorylation of 4 serines within cyclin B1 promotes the rapid nuclear translocation of Cdc2/cyclin B1 at G(2)/M, the role that individual phosphorylation sites play in this process has not been examined (, ). We report here that phosphorylation of a single serine residue (Ser(113) of Xenopus cyclin B1) abrogates nuclear export of cyclin B1. This serine lies directly within the cyclin B1 nuclear export sequence and, when phosphorylated, prevents binding of the nuclear export factor, CRM1. In contrast, analysis of phosphorylation site mutants suggests that coordinate phosphorylation of all 4 serines (94, 96, 101, and 113) is required for the accelerated nuclear import of cyclin B1/Cdc2 characteristic of G(2)/M. Additionally, binding of cyclin B1 to importin-beta, the factor known to be responsible for the slow interphase nuclear entry of cyclin B1, appears to be unaffected by the phosphorylation state of cyclin B. These data suggest that a distinct import factor must be recruited to enhance nuclear entry of Cdc2/cyclin B1 at the G(2)/M transition.
Yang
J
J
Duke University Medical Center, Department of Pharmacology and Cancer Biology, Durham, North Carolina 27710, USA.
Song
H
H
Walsh
S
S
Bardes
E S
ES
Kornbluth
S
S
D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism; D019926:Cyclin B / Q000378:metabolism*; D056744:Cyclin B1; D016195:G2 Phase / Q000502:physiology; D028884:Karyopherins; D008938:Mitosis / Q000502:physiology; D009687:Nuclear Proteins / Q000378:metabolism; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D010766:Phosphorylation; D021382:Protein Sorting Signals / Q000502:physiology; D012694:Serine / Q000378:metabolism*; D014982:Xenopus laevis
D019926:Cyclin B; D056744:Cyclin B1; D028884:Karyopherins; D009687:Nuclear Proteins; D021382:Protein Sorting Signals; D012694:Serine; D016203:CDC2 Protein Kinase