Relocation and distinct subcellular localization of p34cdc2-cyclin B complex at meiosis reinitiation in starfish oocytes.
M phase promoting factor (MPF) is a major element controlling entry into the M phase of the eukaryotic cell cycle. MPF is composed of two subunits, p34cdc2 and cyclin B. Using indirect immunofluorescence staining with specific antibody against starfish cyclin B, we monitored the dynamics of the subcellular distribution of MPF during meiosis reinitiation in starfish oocytes. We found that all of the cyclin B is already associated with p34cdc2 in immature oocytes arrested at the G2/M border and that this inactive complex is present exclusively in the cytoplasm. After its activation, part of the p34cdc2-cyclin B complex moves into the germinal vesicle before nuclear envelope breakdown, independently of either microtubules or actin filaments. Thereafter, some part of the complex accumulates in the nucleolus and condensed chromosomes. Another portion of the complex accumulates on meiotic asters and spindles, while the rest is still present throughout the cytoplasm. As these patterns of localization are detected in the detergent-extracted oocytes, we propose at least four distinct subcellular states of the p34cdc2-cyclin B complex: freely soluble, microtubule-associated, detergent-resistant cytoskeleton-associated and chromosome-associated. Thus, in addition to the intramolecular modification of p34cdc2-cyclin B complex, its intracellular relocation plays a key role in promoting the M phase.
Ookata
K
K
Department of Biological Science, Tokyo Institute of Technology, Yokohama, Japan.
Hisanaga
S
S
Okano
T
T
Tachibana
K
K
Kishimoto
T
T
D000818:Animals; D015153:Blotting, Western; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D016213:Cyclins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D016200:Maturation-Promoting Factor / Q000378:metabolism; D008540:Meiosis; D008856:Microscopy, Fluorescence; D010063:Ovum / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D011233:Precipitin Tests; D006656:Protamine Kinase / Q000378:metabolism; D013215:Starfish
D016213:Cyclins; D006656:Protamine Kinase; D016203:CDC2 Protein Kinase; D016200:Maturation-Promoting Factor