Effects of destruxins, cyclic depsipeptide mycotoxins, on calcium balance and phosphorylation of intracellular proteins in lepidopteran cell lines

Abstract

The influence of the destruxin E, cyclodepsipeptidic mycotoxin produced by the filamentous entomopathogenic fungus Metarhizium anisopliae, on calcium fluxes and protein phosphorylation of in vitro cultivated lepidopteran cell lines have been studied. The use of the fluorescent calcium indicator Fura 2 AM did not show a fast increase in the level of cytosolic calcium in lepidopteran and human cell lines treated with dtx E. In contrast, 45Ca ++ assays detected a late but significant calcium influx in insect cells exposed to dtx E and A for at least 30 min. This effect was not inhibited by calcium channel blockers with the exception of nifedipine 25 μM. A viral treatment potentiated the effect of dtx E on calcium balance. Dtx E also induced a strong, fast (10 min), transient phosphorylation of high molecular weight (250 kDa) cellular proteins. The activation of phosphorylation was only partially inhibited by EDTA. This study provides the first direct evidence of dtx E influence on calcium fluxes and protein phosphorylation.