PMID:15917545

Citation	Leiter, E, Szappanos, H, Oberparleiter, C, Kaiserer, L, Csernoch, L, Pusztahelyi, T, Emri, T, Pócsi, I, Salvenmoser, W and Marx, F (2005) Antifungal protein PAF severely affects the integrity of the plasma membrane of Aspergillus nidulans and induces an apoptosis-like phenotype. Antimicrob. Agents Chemother. 49:2445-53
Abstract	The small, basic, and cysteine-rich antifungal protein PAF is abundantly secreted into the supernatant by the beta-lactam producer Penicillium chrysogenum. PAF inhibits the growth of various important plant and zoopathogenic filamentous fungi. Previous studies revealed the active internalization of the antifungal protein and the induction of multifactorial detrimental effects, which finally resulted in morphological changes and growth inhibition in target fungi. In the present study, we offer detailed insights into the mechanism of action of PAF and give evidence for the induction of a programmed cell death-like phenotype. We proved the hyperpolarization of the plasma membrane in PAF-treated Aspergillus nidulans hyphae by using the aminonaphtylethenylpyridinium dye di-8-ANEPPS. The exposure of phosphatidylserine on the surface of A. nidulans protoplasts by Annexin V staining and the detection of DNA strand breaks by TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) gave evidence for a PAF-induced apoptotic-like mechanism in A. nidulans. The localization of reactive oxygen species (ROS) by dichlorodihydrofluorescein diacetate and the abnormal cellular ultrastructure analyzed by transmission electron microscopy suggested that ROS-elicited membrane damage and the disintegration of mitochondria played a major role in the cytotoxicity of PAF. Finally, the reduced PAF sensitivity of A. nidulans strain FGSC1053, which carries a dominant-interfering mutation in fadA, supported our assumption that G-protein signaling was involved in PAF-mediated toxicity.
Links	PubMed PMC1140496 Online version:10.1128/AAC.49.6.2445-2453.2005
Keywords	Antifungal Agents/pharmacology; Antifungal Agents/toxicity; Apoptosis; Aspergillus nidulans/drug effects; Aspergillus nidulans/ultrastructure; Cell Membrane/drug effects; Cell Membrane/ultrastructure; Cell Membrane Permeability; Fungal Proteins/pharmacology; Fungal Proteins/toxicity; GTP-Binding Proteins/metabolism; Microscopy, Electron, Transmission; Mitochondria/metabolism; Penicillium; Penicillium chrysogenum/metabolism; Phenotype; Reactive Oxygen Species/metabolism; Signal Transduction
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
EMENI:GPA1	involved_in	GO:0035690: cellular response to drug	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete

References

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