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

Overview

Journal Toxins (Basel)

Publisher MDPI

Specialty Toxicology

Date 2017 Apr 6

PMID 28379186

Citations 25

Authors

Slavica Janevska

Birgit Arndt

Leonie Baumann

Lisa Helene Apken

Lucas Maciel Mauriz Marques

Hans-Ulrich Humpf

Bettina Tudzynski

Affiliations

Soon will be listed here.

Abstract

The PKS-NRPS-derived tetramic acid equisetin and its -desmethyl derivative trichosetin exhibit remarkable biological activities against a variety of organisms, including plants and bacteria, e.g., . The equisetin biosynthetic gene cluster was first described in , a species distantly related to the notorious rice pathogen . Here we present the activation and characterization of a homologous, but silent, gene cluster in . Bioinformatic analysis revealed that this cluster does not contain the equisetin -methyltransferase gene and consequently, trichosetin was isolated as final product. The adaption of the inducible, tetracycline-dependent Tet-on promoter system from achieved a controlled overproduction of this toxic metabolite and a functional characterization of each cluster gene in . Overexpression of one of the two cluster-specific transcription factor (TF) genes, , led to an activation of the three biosynthetic cluster genes, including the key gene. In contrast, overexpression of , encoding a second Zn(II)₂Cys₆ TF, did not activate adjacent cluster genes. Instead, was induced by the final product trichosetin and was required for expression of the transporter-encoding gene . TF23 and MFS-T likely act in consort and contribute to detoxification of trichosetin and therefore, self-protection of the producing fungus.

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