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© The Rockefeller University Press, 0021-9525/1999//1261 $5.00
The Journal of Cell Biology, Volume 147, Number 6, , 1999 1261-1274

Original Article

Dynein Intermediate Chain Mediated Dynein–Dynactin Interaction Is Required for Interphase Microtubule Organization and Centrosome Replication and Separation inDictyostelium



Shuo Maa, Leda Triviños-Lagosa, Ralph Gräfb, and Rex L. Chisholma

a Department of Cell and Molecular Biology, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611
b Adolf-Butenandt-Institut/Zellbiologie, Universitaet Muenchen, D-80336 Muenchen, Germany
Department of Cell and Molecular Biology, Northwestern University Medical School, 303 East Chicago Ave., Ward 11-100, Chicago, IL 60611-3072.(312) 503-5994(312) 503-4151

r-chisholm{at}nwu.edu

Cytoplasmic dynein intermediate chain (IC) mediates dynein–dynactin interaction in vitro (Karki, S., and E.L. Holzbaur. 1995. J. Biol. Chem. 270:28806–28811; Vaughan, K.T., and R.B. Vallee. 1995. J. Cell Biol. 131:1507–1516). To investigate the physiological role of IC and dynein–dynactin interaction, we expressed IC truncations in wild-type Dictyostelium cells. IC{Delta}C associated with dynactin but not with dynein heavy chain, whereas IC{Delta}N truncations bound to dynein but bound dynactin poorly. Both mutations resulted in abnormal localization to the Golgi complex, confirming dynein function was disrupted. Striking disorganization of interphase microtubule (MT) networks was observed when mutant expression was induced. In a majority of cells, the MT networks collapsed into large bundles. We also observed cells with multiple cytoplasmic asters and MTs lacking an organizing center. These cells accumulated abnormal DNA content, suggesting a defect in mitosis. Striking defects in centrosome morphology were also observed in IC mutants, mostly larger than normal centrosomes. Ultrastructural analysis of centrosomes in IC mutants showed interphase accumulation of large centrosomes typical of prophase as well as unusually paired centrosomes, suggesting defects in centrosome replication and separation. These results suggest that dynactin-mediated cytoplasmic dynein function is required for the proper organization of interphase MT network as well as centrosome replication and separation in Dictyostelium.

Key Words: dynein function • intermediate chain • dynein–dynactin interaction • microtubule organization • centrosome replication and separation

© 1999 The Rockefeller University Press

. Abbreviations used in this paper: DAPI, 4,6-diamidino-2-phenylindole; HC, heavy chain; IC, intermediate chain; IP, immunoprecipitation; MT, microtubule; MTOC, MT organizing center.


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