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Vol. 10, Issue 1, 225-243, January 1999

Disruption of a Dynamin Homologue Affects Endocytosis, Organelle Morphology, and Cytokinesis in Dictyostelium discoideum

Dirk C. Wienke,* Menno L.W. Knetsch,* Eva M. Neuhaus,dagger Mary C. Reedy,Dagger and Dietmar J. Manstein*§

 *Abteilung Biophysik and  dagger Abteilung Molekulare Zellforschung, Max-Planck-Institut für Medizinische Forschung, D-69120 Heidelberg, Germany; and  Dagger Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710

The identification and functional characterization of Dictyostelium discoideum dynamin A, a protein composed of 853 amino acids that shares up to 44% sequence identity with other dynamin-related proteins, is described. Dynamin A is present during all stages of D. discoideum development and is found predominantly in the cytosolic fraction and in association with endosomal and postlysosomal vacuoles. Overexpression of the protein has no adverse effect on the cells, whereas depletion of dynamin A by gene-targeting techniques leads to multiple and complex phenotypic changes. Cells lacking a functional copy of dymA show alterations of mitochondrial, nuclear, and endosomal morphology and a defect in fluid-phase uptake. They also become multinucleated due to a failure to complete normal cytokinesis. These pleiotropic effects of dynamin A depletion can be rescued by complementation with the cloned gene. Morphological studies using cells producing green fluorescent protein-dynamin A revealed that dynamin A associates with punctate cytoplasmic vesicles. Double labeling with vacuolin, a marker of a postlysosomal compartment in D. discoideum, showed an almost complete colocalization of vacuolin and dynamin A. Our results suggest that that dynamin A is likely to function in membrane trafficking processes along the endo-lysosomal pathway of D. discoideum but not at the plasma membrane.

§   Corresponding author.

Molecular Biology of the Cell
Vol. 10, 225-243, January 1999
Copyright © 1999 by The American Society for Cell Biology


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