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Establishment of the Inducible Tet-On System for the Activation of the Silent Trichosetin Gene Cluster in Fusarium fujikuroi

Institute of Plant Biology and Biotechnology, Westfälische Wilhelms-Universität Münster, Schlossplatz 8, 48143 Münster, Germany
Institute of Food Chemistry, Westfälische Wilhelms-Universität Münster, Corrensstrasse 45, 48149 Münster, Germany
Author to whom correspondence should be addressed.
Academic Editor: Jiujiang Yu
Toxins 2017, 9(4), 126;
Received: 6 February 2017 / Revised: 30 March 2017 / Accepted: 31 March 2017 / Published: 5 April 2017
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The PKS-NRPS-derived tetramic acid equisetin and its N-desmethyl derivative trichosetin exhibit remarkable biological activities against a variety of organisms, including plants and bacteria, e.g., Staphylococcus aureus. The equisetin biosynthetic gene cluster was first described in Fusarium heterosporum, a species distantly related to the notorious rice pathogen Fusarium fujikuroi. Here we present the activation and characterization of a homologous, but silent, gene cluster in F. fujikuroi. Bioinformatic analysis revealed that this cluster does not contain the equisetin N-methyltransferase gene eqxD and consequently, trichosetin was isolated as final product. The adaption of the inducible, tetracycline-dependent Tet-on promoter system from Aspergillus niger achieved a controlled overproduction of this toxic metabolite and a functional characterization of each cluster gene in F. fujikuroi. Overexpression of one of the two cluster-specific transcription factor (TF) genes, TF22, led to an activation of the three biosynthetic cluster genes, including the PKS-NRPS key gene. In contrast, overexpression of TF23, encoding a second Zn(II)2Cys6 TF, did not activate adjacent cluster genes. Instead, TF23 was induced by the final product trichosetin and was required for expression of the transporter-encoding gene MFS-T. TF23 and MFS-T likely act in consort and contribute to detoxification of trichosetin and therefore, self-protection of the producing fungus. View Full-Text
Keywords: Fungi; Fusarium fujikuroi; secondary metabolism; PKS-NRPS; biosynthesis; gene regulation Fungi; Fusarium fujikuroi; secondary metabolism; PKS-NRPS; biosynthesis; gene regulation

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Janevska, S.; Arndt, B.; Baumann, L.; Apken, L.H.; Mauriz Marques, L.M.; Humpf, H.-U.; Tudzynski, B. Establishment of the Inducible Tet-On System for the Activation of the Silent Trichosetin Gene Cluster in Fusarium fujikuroi. Toxins 2017, 9, 126.

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