Systematic Evaluation and Application of IDR Domain-Mediated Transcriptional Activation of NUP98 in Saccharomyces cerevisiae

IF 3.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS ACS Synthetic Biology Pub Date : 2024-10-29 DOI:10.1021/acssynbio.4c0037610.1021/acssynbio.4c00376

Sheng Wang*, Xueming Wu, Zhenghao Qiao, Xuan He, Yu Li, Tianyu Zhang, Weiwei Liu, Ming Wang, Xiangtian Zhou* and Yang Yu*,

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Abstract

Implementing dynamic control over gene transcription to decouple cell growth is essential for regulating protein expression in microbial cells. However, the availability of efficient regulatory elements in Saccharomyces cerevisiae remains limited. In this study, we present a novel β-estradiol-inducible gene expression system, termed DEN. This system combines a DNA-binding domain with an estradiol-binding domain and an intrinsically disordered region (IDR) from NUP98. Comparative analysis shows that the DEN system outperforms IDRs from other proteins, achieving an approximately 60-fold increase in EGFP expression upon β-estradiol induction. Moreover, our system is tightly controlled; nontoxic gene expression makes it a powerful tool for rapid and precise modulation of target gene expression. This system holds great potential for unlocking new functionalities from existing proteins in future research.

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系统评估和应用 IDR 域介导的 NUP98 在酿酒酵母中的转录激活作用

对基因转录实施动态控制，使细胞生长脱钩，对于调节微生物细胞中蛋白质的表达至关重要。然而，在酿酒酵母中可用的高效调控元件仍然有限。在这项研究中，我们提出了一种新型的β-雌二醇诱导基因表达系统，称为DEN。该系统结合了 DNA 结合域、雌二醇结合域和来自 NUP98 的内在无序区（IDR）。比较分析表明，DEN 系统优于其他蛋白质的 IDR，在β-雌二醇诱导下，EGFP 的表达量增加了约 60 倍。此外，我们的系统是严格控制的；无毒基因表达使其成为快速、精确调节靶基因表达的有力工具。在未来的研究中，该系统具有从现有蛋白质中释放新功能的巨大潜力。

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来源期刊

ACS Synthetic Biology 生物-

CiteScore

8.00

自引率

10.60%

发文量

380

审稿时长

6-12 weeks

期刊介绍： The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.