Screening Proteins That Interact With AcHog1 and the Functional Analysis of AcSko1 in Aspergillus cristatus.

IF 3.5 4区 生物学 Q2 MICROBIOLOGY Journal of Basic Microbiology Pub Date : 2024-10-07 DOI:10.1002/jobm.202400475
Lei Shao, Zuoyi Liu, Yongxiang Liu, Yumei Tan
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Abstract

Aspergillus cristatus is a dominant fungus formed during the "flowering" process of Fuzhuan brick tea. Previous research has established that the sporulation of Aspergillus nidulans, a model organism of filamentous fungi, is regulated by light. However, the sporulation of A. cristatus is dependent on osmotic stress. In a previous study, we used pull-down and mass spectrometry to identify proteins that interacted with AcHog1 in A. cristatus when cultured under different conditions of osmotic stress. In the present study, we analyzed the proteins we identified previously to investigate their functional role. The AA1E3BER4 protein was located downstream of Hog1 in the HOG branch pathway and was identified that was regulated by AcHog1. Furthermore, yeast two-hybrid analysis showed that AA1E3BER4 interacted with AcHog1. In addition, we knocked out and complemented the Acsko1 gene encoding the AA1E3BER4 protein. We found that the number of sexual and asexual spores were downregulated by 3.81- and 4.57-fold, respectively, in the ΔAcsko1 strain. The sensitivity of the ΔAcsko1 strain to sorbitol and sucrose, as regulators of osmotic stress, increased, and the sensitivity to high sucrose was higher than that of sorbitol. Acsko1 also regulated the response of A. cristatus to oxidative stress, Congo red, and SDS (sodium dodecyl sulfate). In addition, the deletion of Acsko1 significantly increased the pigment of the ΔAcsko1 strain. This is the first study to report the role of the sko1 gene in oxidative stress, stress-induced damage to the cell wall, and pigment in Aspergillus cristatus.

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筛选与 AcHog1 相互作用的蛋白质并对十字花曲霉中的 AcSko1 进行功能分析
皱缩曲霉是福砖茶 "发花 "过程中形成的优势真菌。先前的研究已经证实,丝状真菌的模式生物黑曲霉的孢子分生受光照调节。然而,冠突散囊菌的孢子生成依赖于渗透胁迫。在之前的一项研究中,我们使用牵引法和质谱法鉴定了在不同渗透胁迫条件下培养十字花科真菌时与 AcHog1 相互作用的蛋白质。在本研究中,我们对之前鉴定出的蛋白质进行了分析,以研究它们的功能作用。AA1E3BER4蛋白位于HOG分支通路中Hog1的下游,并被发现受AcHog1调控。此外,酵母双杂交分析表明,AA1E3BER4 与 AcHog1 相互作用。此外,我们还敲除了编码 AA1E3BER4 蛋白的 Acsko1 基因并对其进行了互补。我们发现,在ΔAcsko1菌株中,有性孢子和无性孢子的数量分别下调了3.81倍和4.57倍。作为渗透胁迫的调节因子,ΔAcsko1菌株对山梨醇和蔗糖的敏感性增加,对高蔗糖的敏感性高于对山梨醇的敏感性。Acsko1 还能调控 A. cristatus 对氧化应激、刚果红和 SDS(十二烷基硫酸钠)的反应。此外,缺失 Acsko1 能显著增加 ΔAcsko1 菌株的色素。这是首次报道 sko1 基因在曲霉氧化应激、应激引起的细胞壁损伤和色素中的作用的研究。
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来源期刊
Journal of Basic Microbiology
Journal of Basic Microbiology 生物-微生物学
CiteScore
6.10
自引率
0.00%
发文量
134
审稿时长
1.8 months
期刊介绍: The Journal of Basic Microbiology (JBM) publishes primary research papers on both procaryotic and eucaryotic microorganisms, including bacteria, archaea, fungi, algae, protozoans, phages, viruses, viroids and prions. Papers published deal with: microbial interactions (pathogenic, mutualistic, environmental), ecology, physiology, genetics and cell biology/development, new methodologies, i.e., new imaging technologies (e.g. video-fluorescence microscopy, modern TEM applications) novel molecular biology methods (e.g. PCR-based gene targeting or cassettes for cloning of GFP constructs).
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