Regulative roles of glutathione reductase and four glutaredoxins in glutathione redox, antioxidant activity, and iron homeostasis of Beauveria bassiana

Appl Microbiol Biotechnol. 2016 Jul;100(13):5907-17. doi: 10.1007/s00253-016-7420-0. Epub 2016 Mar 12.

Abstract

Multiple glutaredoxins (Grx) and glutathione reductase (Glr) are vital for the thiol-disulfide redox system in budding yeast but generally unexplored in filamentous fungi. Here we characterized the Beauveria bassiana redox system comprising dithiol Grx1, monothiol Grx2-4, Grx-like Grx5, and Glr orthologue. Each grx or glr deletion was compensated by increased transcripts of some other grx genes in normal cultures. Particularly, grx3 compensated the absence of grx1, grx2, grx5, or glr under oxidative stress while its absence was compensated only by undeletable grx4 under normal conditions but by most of other undeleted grx and glr genes in response to menadione. Consequently, the redox state was disturbed in Δglr more than in Δgrx3 but not in Δgrx1/2/5. Superoxide dismutases were more active in normal Δgrx1-3 cultures but less in Δgrx5 or Δglr response to menadione. Total catalase activity increased differentially in all the mutant cultures stressed with or without H2O2 while total peroxidase activity decreased more in the normal or H2O2-stressed culture of Δglr than of Δgrx3. Among the mutants, Δgrx3 showed slightly increased sensitivity to menadione or H2O2; Δglr exhibited greater sensitivity to thiol-oxidizing diamide than thiol-reducing 1-chloro-2,4-dinitrobenzene as well as increased sensitivity to the two oxidants. Intriguingly, all the mutants grew slower in a Fe(3+)-inclusive medium perhaps due to elevated transcripts of two Fe(3+) transporter genes. More or fewer phenotypes linked with biocontrol potential were altered in four deletion mutants excluding Δgrx5. All the changes were restored by targeted gene complementation. Overall, Grx3 played more critical role than other Grx homologues in the Glr-dependent redox system of the fungal entomopathogen.

Keywords: Antioxidation; Biological control potential; Entomopathogenic fungi; Gene expression and regulation; Glutaredoxins; Glutathione reductase; Redox homeostasis.

MeSH terms

  • Antioxidants / metabolism*
  • Beauveria / enzymology
  • Beauveria / genetics
  • Beauveria / metabolism*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Glutaredoxins / genetics
  • Glutaredoxins / metabolism*
  • Glutathione / metabolism*
  • Glutathione Reductase / genetics
  • Glutathione Reductase / metabolism*
  • Homeostasis
  • Iron / metabolism*
  • Mutation
  • Oxidation-Reduction
  • Superoxide Dismutase / genetics
  • Superoxide Dismutase / metabolism

Substances

  • Antioxidants
  • Fungal Proteins
  • Glutaredoxins
  • Iron
  • Superoxide Dismutase
  • Glutathione Reductase
  • Glutathione