Identification of a 102 kDa protein (cytocentrin) immunologically related to keratin 19, which is a cytoplasmically derived component of the mitotic spindle pole.
The mAb RK7, previously shown to recognize keratin 19, was also found to cross-react with a biologically unrelated 102 kDa protein, which becomes associated with the poles of the mitotic apparatus. This newly identified protein, called cytocentrin, is a stable cellular component, may be at least in part phosphorylated, and displays a cell cycle-dependent cellular localization. In interphase cells, it is diffusely distributed in the cytosol and shows no affinity for cytoplasmic microtubules. It becomes localized to the centrosome in early prophase, prior to nuclear envelope breakdown, separation of replicated centrosomes, and nucleation of mitotic apparatus microtubules. During metaphase, cytocentrin is located predominately at the mitotic poles, often appearing as an aggregate of small globular sub-components; it also associates with some polar microtubules. In late anaphase/early telophase cytocentrin dissociates entirely from the mitotic apparatus and becomes temporarily localized with microtubules in the midbody, from which it disappears by late telophase. In taxol-treated cells cytocentrin was associated with the center of the miniasters but also showed affinity for some cytoplasmic microtubules. Studies employing G2-synchronized cells and nocodazole demonstrated that cytocentrin can become associated with mitotic centrosomes independently of tubulin polymerization and that microtubules regrow from antigen-containing foci. We interpret these results to suggest that cytocentrin is a cytoplasmic protein that becomes specifically activated or modified at the onset of mitosis so that it can affiliate with the mitotic poles where it may provide a link between the pericentriolar material and other components of the mitotic apparatus.
Paul
E C
EC
Cornell University, Department of Physiology, Ithaca, New York 14853.
Quaroni
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D002453:Cell Cycle; D002460:Cell Line; D003429:Cross Reactions; D003593:Cytoplasm / Q000378:metabolism*; D000939:Epitopes; D007633:Keratins / Q000276:immunology*; D034882:Lamins; D008969:Molecular Sequence Data; D015739:Nocodazole / Q000494:pharmacology; D009687:Nuclear Proteins / Q000032:analysis; D017239:Paclitaxel / Q000494:pharmacology; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D051381:Rats; D008941:Spindle Apparatus / Q000737:chemistry*; D014404:Tubulin / Q000032:analysis
D000911:Antibodies, Monoclonal; D000939:Epitopes; D034882:Lamins; D009687:Nuclear Proteins; D011506:Proteins; D014404:Tubulin; D007633:Keratins; D017239:Paclitaxel; D015739:Nocodazole