Antifungal protein PAF severely affects the integrity of the plasma membrane of Aspergillus nidulans and induces an apoptosis-like phenotype

E. Leiter, Henrietta Szappanos, Christoph Oberparleiter, Lydia Kaiserer, L. Csernoch, T. Pusztahelyi, T. Emri, I. Pócsi, Willibald Salvenmoser, Florentine Marx

Research output: Contribution to journalArticle

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Abstract

The small, basic, and cysteine-rich antifungal protein PAF is abundantly secreted into the supernatant by the β-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.

Original languageEnglish
Pages (from-to)2445-2453
Number of pages9
JournalAntimicrobial Agents and Chemotherapy
Volume49
Issue number6
DOIs
Publication statusPublished - Jun 2005

Fingerprint

Aspergillus nidulans
Cell Membrane
Apoptosis
Phenotype
Reactive Oxygen Species
Proteins
Fungi
Penicillium chrysogenum
Lactams
DNA Breaks
DNA Nucleotidylexotransferase
Hyphae
Protoplasts
Annexin A5
Phosphatidylserines
In Situ Nick-End Labeling
Growth
Biotin
Transmission Electron Microscopy
GTP-Binding Proteins

ASJC Scopus subject areas

  • Pharmacology (medical)

Cite this

Antifungal protein PAF severely affects the integrity of the plasma membrane of Aspergillus nidulans and induces an apoptosis-like phenotype. / Leiter, E.; Szappanos, Henrietta; Oberparleiter, Christoph; Kaiserer, Lydia; Csernoch, L.; Pusztahelyi, T.; Emri, T.; Pócsi, I.; Salvenmoser, Willibald; Marx, Florentine.

In: Antimicrobial Agents and Chemotherapy, Vol. 49, No. 6, 06.2005, p. 2445-2453.

Research output: Contribution to journalArticle

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