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钾离子比钠离子更能促进多胺对基因表达的有利影响。

K+ promotes the favorable effect of polyamine on gene expression better than Na.

机构信息

Faculty of Life and Medical Sciences, Doshisha University, Kyoto, Japan.

Faculty of Science and Engineering, Doshisha University, Kyoto, Japan.

出版信息

PLoS One. 2020 Sep 3;15(9):e0238447. doi: 10.1371/journal.pone.0238447. eCollection 2020.

Abstract

BACKGROUND

Polyamines are involved in a wide variety of biological processes including a marked effect on the structure and function of DNA. During our study on the interaction of polyamines with DNA, we found that K+ enhanced in vitro gene expression in the presence of polyamine more strongly than Na+. Thus, we sought to clarify the physico-chemical mechanism underlying this marked difference between the effects of K+ and Na+.

PRINCIPAL FINDINGS

It was found that K+ enhanced gene expression in the presence of spermidine, SPD(3+), much more strongly than Na+, through in vitro experiments with a Luciferase assay on cell extracts. Single-DNA observation by fluorescence microscopy showed that Na+ prevents the folding transition of DNA into a compact state more strongly than K+. 1H NMR measurement revealed that Na+ inhibits the binding of SPD to DNA more strongly than K+. Thus, SPD binds to DNA more favorably in K+-rich medium than in Na+-rich medium, which leads to favorable conditions for RNA polymerase to access DNA by decreasing the negative charge.

CONCLUSION AND SIGNIFICANCE

We found that Na+ and K+ exhibit markedly different effects through competitive binding with a cationic polyamine, SPD, to DNA, which causes a large difference in the higher-order structure of genomic DNA. It is concluded that the larger favorable effect of Na+ than K+ on in vitro gene expression observed in this study is well attributable to the significant difference between Na+ and K+ on the competitive binding inducing conformational transition of DNA.

摘要

背景

多胺参与多种生物过程,包括对 DNA 结构和功能的显著影响。在研究多胺与 DNA 的相互作用时,我们发现多胺存在时 K+比 Na+更能增强体外基因表达。因此,我们试图阐明 K+和 Na+对基因表达影响的显著差异背后的物理化学机制。

主要发现

通过细胞提取物的荧光素酶测定进行的体外实验发现,K+比 Na+更能增强 spermidine(SPD,3+)存在时的基因表达。荧光显微镜下的单链 DNA 观察表明,Na+比 K+更能强烈地阻止 DNA 折叠成紧凑状态的转变。1H NMR 测量显示,Na+比 K+更能抑制 SPD 与 DNA 的结合。因此,在富含 K+的介质中,SPD 与 DNA 的结合更有利,这通过降低负电荷有利于 RNA 聚合酶与 DNA 接触。

结论和意义

我们发现,Na+和 K+通过与阳离子多胺 SPD 的竞争结合对 DNA 表现出明显不同的影响,这导致基因组 DNA 的高级结构有很大差异。可以得出结论,本研究中观察到的 Na+比 K+更有利于体外基因表达的更大有利影响,很好地归因于 Na+和 K+在诱导 DNA 构象转变的竞争结合上的显著差异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e33a/7470421/d6d8bb072be6/pone.0238447.g001.jpg

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