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用 AtfA/AtfB 介导的 Menadione 应激反应进行全基因组基因表达分析

Genome-Wide Gene Expression Analyses of the AtfA/AtfB-Mediated Menadione Stress Response in .

机构信息

Department of Molecular Biotechnology and Microbiology, Institute of Biotechnology, Faculty of Science and Technology, University of Debrecen, Egyetem tér 1., 4032 Debrecen, Hungary.

ELRN-UD Fungal Stress Biology Research Group, 4032 Debrecen, Hungary.

出版信息

Cells. 2023 Jan 31;12(3):463. doi: 10.3390/cells12030463.

Abstract

The bZIP transcription factors (TFs) govern regulation of development, secondary metabolism, and various stress responses in filamentous fungi. In this work, we carried out genome-wide expression studies employing Illumina RNAseq to understand the roles of the two bZIP transcription factors AtfA and AtfB in . Comparative analyses of transcriptomes of control, Δ, Δ, and ΔΔ mutant strains were performed. Dependence of a gene on AtfA (AtfB) was decided by its differential downregulation both between the reference and Δ (Δ) strains and between the Δ (Δ) and the ΔΔ strains in vegetatively grown cells (mycelia) and asexual spores (conidia) of menadione sodium bisulfite (MSB)-treated or untreated cultures. As AtfA is the primary bZIP TF governing stress-response in , the number of differentially expressed genes for Δ was significantly higher than for Δ in both mycelial and conidial samples, and most of the AtfB-dependent genes showed AtfA dependence, too. Moreover, the low number of genes depending on AtfB but not on AtfA can be a consequence of Δ leading to downregulation of expression. Conidial samples showed much higher abundance of and mRNAs and more AtfA- and AtfB-affected genes than mycelial samples. In the presence of MSB, the number of AtfB- (but not of AtfA-) affected genes decreased markedly, which was accompanied with decreased mRNA levels of in MSB-treated mycelial (reference strain) and conidial (Δ mutant) samples. In mycelia, the overlap between the AtfA-dependent genes in MSB-treated and in untreated samples was low, demonstrating that distinct genes can be under AtfA control under different conditions. Carbohydrate metabolism genes were enriched in the set of AtfA-dependent genes. Among them, AtfA-dependence of glycolytic genes in conidial samples was the most notable. Levels of transcripts of certain secondary metabolitic gene clusters, such as the Emericellamide cluster, also showed AtfA-dependent regulation. Genes encoding catalase and histidine-containing phosphotransfer proteins showed AtfA-dependence under all experimental conditions. There were 23 AtfB-dependent genes that did not depend on AtfA under any of our experimental conditions. These included a putative α-glucosidase (), a putative α-amylase, which is involved in early conidial germination, and an alternative oxidase. In summary, in there is a complex interaction between the two bZIP transcription factors, where AtfA plays the primary regulatory role.

摘要

bZIP 转录因子 (TFs) 调控丝状真菌的发育、次生代谢和各种应激反应。在这项工作中,我们通过 Illumina RNAseq 进行了全基因组表达研究,以了解 AtfA 和 AtfB 这两个 bZIP 转录因子在中的作用。比较了对照、Δ、Δ和 ΔΔ突变菌株的转录组。通过比较参考菌株和 Δ、Δ 和 ΔΔ 菌株之间基因在营养生长细胞(菌丝)和无性孢子(分生孢子)中的差异下调,可以决定一个基因对 AtfA(AtfB)的依赖性。由于 AtfA 是调控 的主要 bZIP TF,因此在菌丝和分生孢子样品中,Δ 的差异表达基因数量明显高于 Δ,并且大多数依赖 AtfB 的基因也依赖 AtfA。此外,依赖 AtfB 但不依赖 AtfA 的基因数量较少可能是由于 Δ 导致 表达下调的结果。与菌丝样品相比,分生孢子样品显示出更高丰度的 和 mRNAs 和更多受 AtfA 和 AtfB 影响的基因。在 MSB 存在下,AtfB-(而非 AtfA-)影响的基因数量显著减少,这伴随着 MSB 处理的菌丝(参考菌株)和分生孢子(Δ 突变体)样品中 mRNA 水平的降低。在菌丝中,MSB 处理和未处理样品中 AtfA 依赖性基因之间的重叠很少,表明在不同条件下可以有不同的基因受到 AtfA 控制。碳水化合物代谢基因在 AtfA 依赖性基因中富集。其中,在分生孢子样品中糖酵解基因对 AtfA 的依赖性最为显著。某些次级代谢基因簇,如 Emericellamide 簇的转录本水平也表现出 AtfA 依赖性调节。在所有实验条件下,编码过氧化氢酶和组氨酸含有磷酸转移蛋白的基因均表现出 AtfA 依赖性。有 23 个 AtfB 依赖性基因在我们的任何实验条件下均不依赖 AtfA。其中包括一个假定的α-葡萄糖苷酶()、一个假定的α-淀粉酶、参与早期分生孢子萌发的 以及替代氧化酶。总之,在 中,这两个 bZIP 转录因子之间存在复杂的相互作用,其中 AtfA 发挥主要调节作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3308/9913763/b1ed2394440b/cells-12-00463-g001.jpg

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