Papers in the Biological Sciences


Date of this Version

September 2000


Published in Published in Abstracts: 40th American Society for Cell Biology Annual Meeting, in Molecular Biology of the Cell 11 (supplement): p. 327a. Copyright © 2000 American Society for Cell Biology. Used by permission. Used by permission.


The Ca2+/calmodul1n-dependent protein phosphatase calcinewin is involved in the development of the cellular slime mold Dictyostelium discoideum. Because of its interactions with Ca2+, which appear to influence D. discoideum phagocytosis (Yuan and Chia, 1999, Mol. Biol. Cell 10, 220a), we undertook studies to test whether calcineurin also plays a role in Dictyostelium phagocytosis. The immunosuppressants cyclosporin A and FK506, through the formation of cyclosporin A-cyclophilin A and FK506- FK506-binding protein complexes, respectively, inhibited calcineurin activity. These two calcineurin inhibitors suppressed phagocytosis of fluorescently labeled yeast in a dose-dependent manner. Although it inhib~ted phagocytosis, cyclosporin A had an insignificant effect on the macropinocytosis of the fluid-phase marker fluorescein isothiocyanatedextran. Furthermore, trifluoperazine, a calmodulin antagonist that indirectly inhibits calcinewin, also suppressed phagocytosis in a dosedependent fashion and induced the formation of giant intracellular vacuoles Fluorescence microscopy of cyclosporin A-treated (for 30 min.) cells stained with rhodamine-phalloidin had cytoplasmic chunks of F-actin that were not present in control cells, while cells treated with FK506 and trifluoperazine (also for 30 min.), displayed less cortical but more cytoplasmic F-actin staining than normal cells. Typically, drug-treated cells were smaller and rounder than untreated cells. Our data suggest calcineurin may play a role in D. discoideum phagocytosis, either through the dephosphorylation of actinregulating proteins or other cytoskeletal proteins such as the heavy chain subunit of nonmuscle myosin I1 since dephosphorylation of the latter promotes filament assembly.

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