大分子拥挤对转录及其调控的影响。

Q3 Biochemistry, Genetics and Molecular Biology
QRB Discovery Pub Date : 2025-04-03 eCollection Date: 2025-01-01 DOI:10.1017/qrd.2025.8
Wenxuan Xu, Dylan Collette, Jin Qian, Laura Finzi, David Dunlap
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引用次数: 0

摘要

DNA转录成RNA是生命赖以生存的基本细胞过程。它以几种不同的方式受到严格调节,其中最重要的机制是蛋白质诱导的DNA拓扑变化,如环。在体内,无论是转录,还是单个分子所表现出的蛋白质诱导的环动力学都不容易监测。体外单分子方法确实提供了这种可能性,但检测是在稀薄的盐水缓冲条件下进行的,这与拥挤的细胞内环境有很大不同。在下文中,我们描述了在不同浓度的拥挤物存在下监测单个DNA分子的转录和lac抑制因子介导的DNA环,以弥合体外和体内实验之间的差距。我们发现拥挤改变了环状复合物中DNA链的首选方向。拥挤也会降低转录物的延伸率,提高末端的读透率。显然,参与基因调控的蛋白质的活动被拥挤以令人惊讶的方式改变了。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Insights on the effect of macromolecular crowding on transcription and its regulation.

Transcription of DNA into RNA is a fundamental cellular process upon which life depends. It is tightly regulated in several different ways, and among the most important mechanisms are protein-induced topological changes in DNA such as looping. In vivo neither transcription, nor protein-induced looping dynamics exhibited by individual molecules are easily monitored. In vitro single-molecule approaches do offer that possibility, but assays are conducted in rarefied, saline buffer conditions which greatly differ from the crowded intracellular environment. In the following, we describe monitoring both transcription and lac repressor-mediated DNA looping of single DNA molecules in the presence of different concentrations of crowders to bridge the gap between in vitro and in vivo experimentation. We found that crowding shifts the preferred orientation of DNA strands in the looped complex. Crowding also attenuates the rate of transcript elongation and enhances readthrough at the terminator. Clearly, the activities of proteins involved in gene regulation are modified in surprising ways by crowding.

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来源期刊
QRB Discovery
QRB Discovery Biochemistry, Genetics and Molecular Biology-Biophysics
CiteScore
3.60
自引率
0.00%
发文量
18
审稿时长
12 weeks
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