Dynein intermediate chain mediated dynein-dynactin interaction is required for interphase microtubule organization and centrosome replication and separation in Dictyostelium.
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. ICDeltaC associated with dynactin but not with dynein heavy chain, whereas ICDeltaN 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.
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Department of Cell and Molecular Biology, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Triviños-Lagos
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Gräf
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Chisholm
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D000818:Animals; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D048429:Cell Size; D018385:Centrosome / Q000378:metabolism* / Q000648:ultrastructure; D020090:Chromosome Segregation; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D004023:Dictyostelium / Q000166:cytology* / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D000072159:Dynactin Complex; D004398:Dyneins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D020043:Genes, Essential / Q000235:genetics / Q000502:physiology; D006056:Golgi Apparatus / Q000378:metabolism; D007399:Interphase; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D008938:Mitosis / Q000235:genetics; D008954:Models, Biological; D010641:Phenotype; D011485:Protein Binding; D017384:Sequence Deletion; D013997:Time Factors
D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; D004247:DNA; D004398:Dyneins