J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/09/1055/12 $2.00
Volume 138, Number 5, September 8, 1997 1055-1066

Mitotic Spindle Poles are Organized by Structural and Motor Proteins in Addition to Centrosomes

Tirso Gaglio, Mary A. Dionne, and Duane A. Compton

Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755

The focusing of microtubules into mitotic spindle poles in vertebrate somatic cells has been assumed to be the consequence of their nucleation from centrosomes. Contrary to this simple view, in this article we show that an antibody recognizing the light intermediate chain of cytoplasmic dynein (70.1) disrupts both the focused organization of microtubule minus ends and the localization of the nuclear mitotic apparatus protein at spindle poles when injected into cultured cells during metaphase, despite the presence of centrosomes. Examination of the effects of this dynein-specific antibody both in vitro using a cell-free system for mitotic aster assembly and in vivo after injection into cultured cells reveals that in addition to its direct effect on cytoplasmic dynein this antibody reduces the efficiency with which dynactin associates with microtubules, indicating that the antibody perturbs the cooperative binding of dynein and dynactin to microtubules during spindle/aster assembly. These results indicate that microtubule minus ends are focused into spindle poles in vertebrate somatic cells through a mechanism that involves contributions from both centrosomes and structural and microtubule motor proteins. Furthermore, these findings, together with the recent observation that cytoplasmic dynein is required for the formation and maintenance of acentrosomal spindle poles in extracts prepared from Xenopus eggs (Heald, R., R. Tournebize, T. Blank, R. Sandaltzopoulos, P. Becker, A. Hyman, and E. Karsenti. 1996. Nature (Lond.). 382: 420-425) demonstrate that there is a common mechanism for focusing free microtubule minus ends in both centrosomal and acentrosomal spindles. We discuss these observations in the context of a search-capture-focus model for spindle assembly.

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• Zatsepina, O., Rousselet, A, Chan, P., Olson, M., Jordan, E., Bornens, M (1999). The nucleolar phosphoprotein B23 redistributes in part to the spindle poles during mitosis. J. Cell Sci. 112: 455-466 [Abstract]  
• Bobinnec, Y., Khodjakov, A., Mir, L.M., Rieder, C.L., Edde, B., Bornens, M. (1998). Centriole Disassembly In Vivo and Its Effect on Centrosome Structure and Function in Vertebrate Cells. JCB 143: 1575-1589 [Abstract] [Full Text]  
• Schumacher, J. M., Golden, A., Donovan, P. J. (1998). AIR-2: An Aurora/Ipl1-related Protein Kinase Associated with Chromosomes and Midbody Microtubules Is Required for Polar Body Extrusion and Cytokinesis in Caenorhabditis elegans Embryos. JCB 143: 1635-1646 [Abstract] [Full Text]  
• Wittmann, T., Boleti, H., Antony, C., Karsenti, E., Vernos, I. (1998). Localization of the Kinesin-like Protein Xklp2 to Spindle Poles Requires a Leucine Zipper, a Microtubule-associated Protein, and Dynein. JCB 143: 673-685 [Abstract] [Full Text]  
• Klein, C., Cheutin, T., O'Donohue, M.-F., Rothblum, L., Kaplan, H., Beorchia, A., Lucas, L., Héliot, L., Ploton, D. (1998). The Three-dimensional Study of Chromosomes and Upstream Binding Factor-immunolabeled Nucleolar Organizer Regions Demonstrates Their Nonrandom Spatial Arrangement during Mitosis. Mol. Biol. Cell 9: 3147-3159 [Abstract] [Full Text]  
• Bonaccorsi, S., Giansanti, M. G., Gatti, M. (1998). Spindle Self-organization and Cytokinesis During Male Meiosis in asterless Mutants of Drosophila melanogaster. JCB 142: 751-761 [Abstract] [Full Text]  
• Dujardin, D., Wacker, U. I., Moreau, A., Schroer, T. A., Rickard, J. E., De Mey, J. R. (1998). Evidence for a Role of CLIP-170 in the Establishment of Metaphase Chromosome Alignment. JCB 141: 849-862 [Abstract] [Full Text]  
• Whitehead, C., Rattner, J. (1998). Expanding the role of HsEg5 within the mitotic and post-mitotic phases of the cell cycle. J. Cell Sci. 111: 2551-2561 [Abstract]  
• Inoue, S, Yoder, O., Turgeon, B., Aist, J. (1998). A cytoplasmic dynein required for mitotic aster formation in vivo. J. Cell Sci. 111: 2607-2614 [Abstract]  
• Compton, D. (1998). Focusing on spindle poles. J. Cell Sci. 111: 1477-1481 [Abstract]  
• Inoue, S, Turgeon, B., Yoder, O., Aist, J. (1998). Role of fungal dynein in hyphal growth, microtubule organization, spindle pole body motility and nuclear migration. J. Cell Sci. 111: 1555-1566 [Abstract]  
• Merdes, A., Cleveland, D. W. (1997). Pathways of Spindle Pole Formation: Different Mechanisms; Conserved Components. JCB 138: 953-956 [Full Text]  
• Vorobjev, I., Malikov, V., Rodionov, V. (2001). Self-organization of a radial microtubule array by dynein-dependent nucleation of microtubules. Proc. Natl. Acad. Sci. USA 98: 10160-10165 [Abstract] [Full Text]  
• Howell, B.J., McEwen, B.F., Canman, J.C., Hoffman, D.B., Farrar, E.M., Rieder, C.L., Salmon, E.D. (2001). Cytoplasmic dynein/dynactin drives kinetochore protein transport to the spindle poles and has a role in mitotic spindle checkpoint inactivation. JCB 155: 1159-1172 [Abstract] [Full Text]  

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