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Yeast two-hybrid screening reveals a dual function for the histone acetyltransferase GcnE by controlling glutamine synthesis and development in Aspergillus fumigatus

Abstract

The acetyltransferase GcnE is part of the SAGA complex which regulates fungal gene expression through acetylation of chromatin. Target genes of the histone acetyltransferase GcnE include those involved in secondary metabolism and asexual development. Here, we show that the absence of GcnE not only abrogated conidiation, but also strongly impeded vegetative growth of hyphae in the human pathogenic fungus Aspergillus fumigatus. A yeast two-hybrid screen using a Saccharomyces cerevisiae strain whose tRNA molecules were specifically adapted to express A. fumigatus proteins identified two unprecedented proteins that directly interact with GcnE. Glutamine synthetase GlnA as well as a hypothetical protein located on chromosome 8 (GbpA) were identified as binding partners of GcnE and their interaction was confirmed in vivo via bimolecular fluorescence complementation. Phenotypic characterization of gbpA and glnA deletion mutants revealed a role for GbpA during conidiogenesis and confirmed the central role of GlnA in glutamine biosynthesis. The increase of glutamine synthetase activity in the absence of GcnE indicated that GcnE silences GlnA through binding. This finding suggests an expansion of the regulatory role of GcnE in A. fumigatus.

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Acknowledgements

Lisa Reimer is acknowledged for excellent technical assistance. This study was supported by the Ernst-Abbe University of Applied Sciences Jena, by the Leibniz Institute for Natural Product Research and Infection Biology-Hans Knöll Institute Jena and the collaborative research center/transregio 124 FungiNet (Project A1 and Z2) funded by the Deutsche Forschungsgemeinschaft.

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Correspondence to Falk Hillmann, Thomas Munder or Axel A. Brakhage.

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Communicated by M. Kupiec.

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Nossmann, M., Boysen, J.M., Krüger, T. et al. Yeast two-hybrid screening reveals a dual function for the histone acetyltransferase GcnE by controlling glutamine synthesis and development in Aspergillus fumigatus. Curr Genet 65, 523–538 (2019). https://doi.org/10.1007/s00294-018-0891-z

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Keywords

  • Aspergillus fumigatus
  • Yeast two-hybrid system
  • Histone acetyltransferase GcnE
  • Glutamine synthetase GlnA
  • Bimolecular fluorescence complementation assay