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隔离子网络复合物的一个组成部分,AaSepM,参与了粗糙脉孢菌中多个纤维素响应信号通路。

A component of the septation initiation network complex, AaSepM, is involved in multiple cellulose-responsive signaling pathways in Aspergillus aculeatus.

机构信息

Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, 599-8531, Japan.

Graduate School of Bioresources, Mie University, 1577 Kurimamachiya-cho, Tsu, 514-8507, Japan.

出版信息

Appl Microbiol Biotechnol. 2021 Feb;105(4):1535-1546. doi: 10.1007/s00253-021-11110-7. Epub 2021 Jan 22.

Abstract

Various carbohydrate-active enzymes in Aspergillus are produced in response to physiological inducers, which is regulated at the transcriptional level. To elucidate the induction mechanisms in Aspergillus, we screened for new regulators involved in cellulose-responsive induction from approximately 10,000 Aspergillus aculeatus T-DNA-inserted mutants. We constructed the T-DNA-inserted mutant library using the host strain harboring the orotidine 5'-monophosphate decarboxylase gene (pyrG) under the control of the FIII-avicelase gene (cbhI) promoter. Thus, candidate mutants deficient in cellulose-responsive induction were positively screened via counter selection against 5-fluoroorotic acid (5-FOA). Among less than two hundred 5-FOA-resistant mutants, one mutant that the T-DNA inserted into the AasepM locus reduced the cbhI expression in response to cellulose. Since AaSepM is similar to Schizosaccharomyces pombe Cdc14p (E-value, 2e-20; identities, 33%), which is a component of the septation initiation network (SIN)-complex, we constructed an AasepM deletion mutant (ΔAasepM). We analyzed the expression of cellulase and xylanase genes in response to cellulose, septation, and conidiation in ΔAasepM. The AasepM deletion leads to delayed septation and decreased formation of the conidium chain in A. aculeatus but does not affect hyphal growth on minimal media. We also confirmed AaSepM's involvement in multiple cellulose-responsive signaling pathways of cellulase and xylanase genes under the control of the ManR-dependent, XlnR-dependent, and ManR- and XlnR-independent signaling pathways. KEY POINTS : • A new regulator for cellulolytic gene expression has been identified. • AaSepM is involved in septation and conidiation in A. aculeatus. • AasepM is involved in multiple cellulose-responsive signaling pathways.

摘要

各种曲霉中的碳水化合物活性酶是对生理诱导物作出反应而产生的,这种反应受转录水平调控。为了阐明曲霉中的诱导机制,我们从大约 10000 个曲霉刺座枝孢霉 T-DNA 插入突变体中筛选了新的参与纤维素响应诱导的调控因子。我们使用宿主菌株构建了 T-DNA 插入突变体文库,该菌株中尿嘧啶 5′-单磷酸脱羧酶基因(pyrG)受 FIII-内切纤维素酶基因(cbhI)启动子的控制。因此,通过对 5-氟乳清酸(5-FOA)的反选择,可正向筛选纤维素响应诱导缺陷的候选突变体。在不到 200 个 5-FOA 抗性突变体中,一个 T-DNA 插入到 AasepM 基因座的突变体,其 cbhI 表达对纤维素的响应降低。由于 AaSepM 与裂殖酵母 Schizosaccharomyces pombe Cdc14p(E 值,2e-20;同一性,33%)相似,后者是有丝分裂起始网络(SIN)-复合物的一个组成部分,我们构建了 AasepM 缺失突变体(ΔAasepM)。我们分析了 ΔAasepM 中纤维素酶和木聚糖酶基因对纤维素、有丝分裂和分生孢子形成的响应。AasepM 缺失导致刺座枝孢霉有丝分裂延迟和分生孢子链形成减少,但不影响菌丝在最低培养基上的生长。我们还证实 AaSepM 参与了受 ManR 依赖、XlnR 依赖和 ManR 和 XlnR 非依赖信号通路控制的多个纤维素响应的纤维素酶和木聚糖酶基因的信号通路。要点:• 鉴定了一个新的纤维素表达调控因子。• AaSepM 参与了刺座枝孢霉的有丝分裂和分生孢子形成。• AasepM 参与了多个纤维素响应信号通路。

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