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Global gene regulation by RNA-binding protein NrdA in Aspergillus luchuensis mut. kawachii

ABSTRACT: We report that the RNA-binding protein NrdA has a significant role on the global mRNA expression in the white koji fungus, Aspergillus luchuensis mut. kawachii. We constructed the nrdA conditional expression strain using the Tet-On system in A. kawachii. Downregulation of nrdA caused severe growth defect, indicating that NrdA is essential for proliferation of A. kawachii. Parallel RNA-seq and RNA immunoprecipitation (RIP)-seq analysis identified potential NrdA-interacted transcripts, corresponding to 32% of predicted protein coding genes of A. kawachii. Subsequent gene ontology analysis suggested that overexpression of NrdA affects a variety of metabolic activity including secondary metabolite production.

ORGANISM(S): Aspergillus luchuensis

PROVIDER: GSE164392 | GEO | 2021/01/08

REPOSITORIES: GEO

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Project description:A koji mold Aspergillus kawachii is used for making a Japanese distilled spirit, shochu. During making shochu, A. kawachii is grown in a solid-state culture comprised of steamed grains such as rice or barley, termed koji, to convert the starch to glucose and also to produce citric acid. During this process, cultivation temperature of A. kawachii is generally controlled by raising to 40M-BM-0C and then lowering to 30M-BM-0C. The cultivation at 40M-BM-0C and the following at lower temperature of 30M-BM-0C has important roles in the elevated activity of amylase and starting an accumulation of a large amount of citric acid, respectively. In this study, we investigated the effect of temperature on the gene expression of A. kawachii when barley was used as ingredient for making koji. The results of DNA microarray and gene ontology analysis showed that the expression of genes involved in the metabolic processes of glycerol, trehalose, and pentose phosphate that branch from glycolysis was down-regulated by shifting the cultivation temperature from 40M-BM-0C from 30M-BM-0C. In addition, reduction in the expression of the genes related with heat shock responses and increasing in the expression of the genes related with amino acid transport after the temperature lowering were found to be significant change. These results suggested that the cultivation at 40M-BM-0C is stressful event for the A. kawachii and the heat adaption lead to the depression of citric acid accumulation through activation of the branching pathways from glycolysis. The gene expression profile obtained in this study will help to understand the gene regulation during koji-making process to optimize A. kawachii as industrial microorganism. Gene expression of Aspergillus kawachii in barley koji was measured at 25, 26.5, and 44 hours at two different temperature control. Three independent experiments were performed at each time (25, 26.5, or 44 hours) using different barley koji for each expreriment.

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Global gene regulation by RNA-binding protein NrdA in Aspergillus luchuensis mut. kawachii

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