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[酵母中tRNA摆动碱基修饰与硼酸抗性:抗硼缺失突变体诱导一般氨基酸控制机制并激活硼外流]

[tRNA Wobble Base Modifications and Boric Acid Resistance in Yeast: Boron-Resistant Deletion Mutants Induce the General Amino Acid Control Mechanism and Activate Boron Efflux].

作者信息

Uluisik I, С Karakaya H, Koc A

机构信息

Department of Molecular Biology and Genetics, Izmir Institute of Technology, Izmir, 35430 Turkey.

Department of Biomedical Engineering, Iskenderun Technical University, Hatay, 31200 Turkey.

出版信息

Mol Biol (Mosk). 2020 May-Jun;54(3):450-456. doi: 10.31857/S0026898420030180.

Abstract

Boric acid is essential for plants and has many vital roles in animals and microorganisms. However, its high doses are toxic to all organisms. We previously screened yeast deletion collections to identify boric acid-resistant and susceptible mutants to identify genes that play a role in boron tolerance. Here, we analyzed boron resistant mutants (elplΔ, elp3Δ, elp6Δ, ncs2Δ, ncs6Δ and ktil2Δ) for their abilities to modulate the general amino acid control system (GAAC) and to induce boron efflux pump ATR1. The mutants analyzed in this study lack the genes that play roles in tRNA Wobble base modifications. We found that all of the boron resistant mutants activated Gcn4-dependent reporter gene activity and increased the transcript level of the ATR1 gene. Additionally, boron resistant cells accumulated less boric acid in their cytoplasm compared to the wild type cells upon boron exposure. Thus, our findings suggested that loss of wobble base modifications in tRNA leads to GAAC activation and ATR1 induction, which in turn reduced intracellular boron levels and caused boron resistance.

摘要

硼酸对植物至关重要,在动物和微生物中也有许多重要作用。然而,高剂量的硼酸对所有生物都有毒性。我们之前筛选了酵母缺失文库,以鉴定对硼酸具有抗性和敏感性的突变体,从而确定在硼耐受性中起作用的基因。在此,我们分析了硼抗性突变体(elplΔ、elp3Δ、elp6Δ、ncs2Δ、ncs6Δ和ktil2Δ)调节一般氨基酸控制系统(GAAC)和诱导硼外流泵ATR1的能力。本研究中分析的突变体缺乏在tRNA摆动碱基修饰中起作用的基因。我们发现,所有硼抗性突变体均激活了Gcn4依赖性报告基因活性,并提高了ATR1基因的转录水平。此外,与野生型细胞相比,硼抗性细胞在暴露于硼时,其细胞质中积累的硼酸较少。因此,我们的研究结果表明,tRNA中摆动碱基修饰的缺失导致GAAC激活和ATR1诱导,进而降低细胞内硼水平并产生硼抗性。

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