Regulation of pseurotin A biosynthesis by GliZ and the synergetic effects of gliotoxin and pseurotin A biosynthesis on virulence of Aspergillus fumigatus

Abstract Recently, we reported that zinc regulates gliotoxin biosynthesis via ZafA, which is a zinc-responsive transcriptional activator. From an HPLC analysis of culture media of Aspergillus fumigatus, we found a trend of decreasing gliotoxin production but increasing pseurotin A and fumagillin production in proportion to the zinc concentration. The expression of the genes involved in pseurotin A biosynthesis was upregulated under high zinc concentrations. Furthermore, upregulated expression of pseurotin A biosynthetic genes and higher production of pseurotin A were observed in the ZafA deletion strain. Interestingly, the deletion of GliZ, a transcriptional activator of gliotoxin biosynthesis genes, resulted in upregulated expression of pseurotin A biosynthetic genes and increased production of pseurotin A. We detected upregulation of FumR expression in the GliZ and ZafA deletion mutants. The overexpression of GliZ observed in the ZafA deletion mutant resulted in the failure of the mutant to increase pseurotin A production, which is a phenotype of the ZafA deletion mutant. These results suggest that ZafA sequentially regulates pseurotin A biosynthesis through GliZ. Finally, we found through a murine virulence test that the GliZ and FumR double-deletion mutants showed a delayed death rate compared with the single-deletion mutants of either GliZ or FumR. Taken together, these results suggested that the biosyntheses of gliotoxin and pseurotin A are regulated in opposite ways by zinc utilization and that each secondary metabolite is synthesized when the synthesis of another secondary metabolite fails to protect it against the defense system of the host.