4.6 Article

Functional and transcriptomic analysis of the key unfolded protein response transcription factor HacA in Aspergillus oryzae

Journal

GENE
Volume 593, Issue 1, Pages 143-153

Publisher

ELSEVIER
DOI: 10.1016/j.gene.2016.08.018

Keywords

Aspergillus oryzae; HacA; Protein secretion; Repression under secretion stress; RNA-seq; Unfolded protein response

Funding

  1. State 863 Project [2014AA021304]
  2. Science and Technology Planning Project of Guangdong Province [2013B010404007, 2013B090800003, 2016A050503016]
  3. Fundamental Research Funds for the Central Universities [2015ZP032, 2015ZZ040]
  4. Science and Technology Planning Project of Guangzhou City [201510010191]

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HacA is a conserved basic leucine zipper transcription factor that serves as the master transcriptional regulator in the unfolded protein response (UPR). To comprehensively evaluate the role of HacA in Aspergillus oryzae, a homokaryotic hacA disruption mutant (HacA-DE) and a strain that expressed a constitutively active form of HacA (HacA-CA) were successfully generated, and transcriptome analyses of these mutants were performed. Growth and phenotypic profiles demonstrated that hyphal growth and sporulation were impaired in the HacA-DE and HacA-CA strains that were grown on complete and minimal media, and the growth impairment was more pronounced for the HacA-CA strain. Compared with a wild-type (WO strain, the transcriptome results indicated that differentially expressed genes in these mutants mainly fell into four categories: the protein secretory pathway, amino acid metabolism, lipid metabolism, and carbohydrate metabolism. Furthermore, we identified 80 and 36 genes of the secretory pathway whose expression significantly differed in the HacA-CA strain (compared with the WT and HacA-DE strains) and HacA-DE strain (compared with the WT strain), respectively, which mostly belonged to protein folding/UPR, glycosylation, and vesicle transport processes. Both the HacA-CA and HacA-DE strains exhibited reduced expression of extracellular enzymes, especially amylolytic enzymes, which resulted from the activation of the repression under secretion stress mechanism in response to endoplasmic reticulum stress. Collectively, our results suggest that the function of HacA is important not only for UPR induction, but also for growth and fungal physiology, as it serves to reduce secretion stress in A. oryzae. (C) 2016 Elsevier B.V. All rights reserved.

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