Cytoplasmic accumulation of cyclin B1 in human cells: association with a detergent-resistant compartment and with the centrosome.
Mitotic cyclins are thought to function as key regulatory subunits of the universal M-phase-promoting p34cdc2 protein kinase. Previous immunolocalization studies have demonstrated that a fraction of p34cdc2 undergoes cell cycle-dependent accumulation at the centrosome during G2/M. In order to identify the mitotic cyclins with which this p34cdc2 fraction interacts, we carefully examined the subcellular distribution of both cyclin A and B1 in HeLa cells. We show here that part of cyclin B1 is associated with duplicating centrosomes throughout its accumulation in the cytoplasm and up to metaphase. In contrast cyclin A does not exhibit centrosomal association except at the onset of mitosis, from preprophase up to metaphase. We also present cytological and biochemical evidence that cyclin B1 is preferentially accumulated as a detergent-insoluble protein independently of the state of microtubule assembly and under experimental conditions where most of p34cdc2 is soluble. Interestingly, the electrophoretic pattern of the minor insoluble p34cdc2 fraction was previously shown to be particularly enriched in slow-migrating and presumably hyperphosphorylated isoforms, known to interact specifically with cyclin B1 during interphase. From these results we propose that the interaction of cyclin B1 with the centrosomes and with the cytoplasmic structures is a constitutive feature of the mechanism whereby a fraction of p34cdc2 is recruited and subsequently targeted to the cyclin B-dependent activation pathway.
Bailly
E
E
Centre de Génétique Moléculaire-CNRS, Gif sur Yvette, France.
Pines
J
J
Hunter
T
T
Bornens
M
M
D015153:Blotting, Western; D002451:Cell Compartmentation; D002478:Cells, Cultured; D002875:Chromosomes / Q000378:metabolism*; D016213:Cyclins / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D003902:Detergents; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D017830:Octoxynol; D011092:Polyethylene Glycols; D016196:S Phase; D012492:Salts
D016213:Cyclins; D003902:Detergents; D012492:Salts; D011092:Polyethylene Glycols; D017830:Octoxynol