Use of the Aspergillus oryzae actin gene promoter in a novel reporter system for exploring antifungal compounds and their target genes

Junichiro Marui, Akira Yoshimi, Daisuke Hagiwara, Yoshimi Fujii-Watanabe, Ken Oda, Hideaki Koike, Koichi Tamano, Tomoko Ishii, Motoaki Sano, Masayuki MacHida, Keietsu Abe

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

Demand for novel antifungal drugs for medical and agricultural uses has been increasing because of the diversity of pathogenic fungi and the emergence of drug-resistant strains. Genomic resources for various living species, including pathogenic fungi, can be utilized to develop novel and effective antifungal compounds. We used Aspergillus oryzae as a model to construct a reporter system for exploring novel antifungal compounds and their target genes. The comprehensive gene expression analysis showed that the actin-encoding actB gene was transcriptionally highly induced by benomyl treatment. We therefore used the actB gene to construct a novel reporter system for monitoring responses to cytoskeletal stress in A. oryzae by introducing the actB promoter::EGFP fusion gene. Distinct fluorescence was observed in the reporter strain with minimum background noise in response to not only benomyl but also compounds inhibiting lipid metabolism that is closely related to cell membrane integrity. The fluorescent responses indicated that the reporter strain can be used to screen for lead compounds affecting fungal microtubule and cell membrane integrity, both of which are attractive antifungal targets. Furthermore, the reporter strain was shown to be technically applicable for identifying novel target genes of antifungal drugs triggering perturbation of fungal microtubules or membrane integrity.

Original languageEnglish
Pages (from-to)1829-1840
Number of pages12
JournalApplied Microbiology and Biotechnology
Volume87
Issue number5
DOIs
Publication statusPublished - 2010 Aug

Keywords

  • Actin
  • Antifungal
  • Aspergillus oryzae
  • Cytoskeleton
  • Reporter

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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