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Applied and Environmental Microbiology, April 2007, p. 2128-2134, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.02497-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The Antifungal Protein AFP from Aspergillus giganteus Inhibits Chitin Synthesis in Sensitive Fungi{triangledown}

Silke Hagen,1 Florentine Marx,3 Arthur F. Ram,2 and Vera Meyer1,2*

Berlin University of Technology, Institute of Biotechnology, Department Microbiology and Genetics, Berlin, Germany,1 Leiden University, Institute of Biology, Fungal Genetics Research Group, Leiden, The Netherlands,2 Innsbruck Medical University Biocenter, Division of Molecular Biology, Innsbruck, Austria3

Received 25 October 2006/ Accepted 21 January 2007

The antifungal protein AFP from Aspergillus giganteus is highly effective in restricting the growth of major human- and plant-pathogenic filamentous fungi. However, a fundamental prerequisite for the use of AFP as an antifungal drug is a complete understanding of its mode of action. In this study, we performed several analyses focusing on the assumption that the chitin biosynthesis of sensitive fungi is targeted by AFP. Here we show that the N-terminal domain of AFP (amino acids 1 to 33) is sufficient for efficient binding of AFP to chitin but is not adequate for inhibition of the growth of sensitive fungi. AFP susceptibility tests and SYTOX Green uptake experiments with class III and class V chitin synthase mutants of Fusarium oxysporum and Aspergillus oryzae showed that deletions made the fungi less sensitive to AFP and its membrane permeabilization effect. In situ chitin synthase activity assays revealed that chitin synthesis is specifically inhibited by AFP in sensitive fungi, indicating that AFP causes cell wall stress and disturbs cell integrity. Further evidence that there was AFP-induced cell wall stress was obtained by using an Aspergillus niger reporter strain in which the cell wall integrity pathway was strongly induced by AFP.

* Corresponding author. Mailing address: Berlin University of Technology, Institute of Biotechnology, Department Microbiology and Genetics, Gustav-Meyer-Allee 25, D-13355 Berlin, Germany. Phone: 49 30 31472827. Fax: 49 30 31472922. E-mail: v.meyer{at}lb.tu-berlin.de.

{triangledown} Published ahead of print on 2 February 2007.

Applied and Environmental Microbiology, April 2007, p. 2128-2134, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.02497-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Meyer, V., Damveld, R. A., Arentshorst, M., Stahl, U., van den Hondel, C. A. M. J. J., Ram, A. F. J. (2007). Survival in the Presence of Antifungals: GENOME-WIDE EXPRESSION PROFILING OF ASPERGILLUS NIGER IN RESPONSE TO SUBLETHAL CONCENTRATIONS OF CASPOFUNGIN AND FENPROPIMORPH. J. Biol. Chem. 282: 32935-32948 [Abstract] [Full Text]  


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