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温度诱导的狭窄 DNA 弯曲范围维持了细菌启动子在体外的最大活性。

A temperature-induced narrow DNA curvature range sustains the maximum activity of a bacterial promoter in vitro.

机构信息

Istituto Pasteur Fondazione Cenci Bolognetti, Dip. Biologia Cellulare e dello Sviluppo, Sapienza Univ. Roma, via dei Sardi 70, 00185 Roma, Italy.

出版信息

Biochemistry. 2010 Apr 6;49(13):2778-85. doi: 10.1021/bi902003g.

Abstract

Among the molecular strategies bacteria have set up to quickly match their transcriptional program to new environments, changes in sequence-mediated DNA curvature play a crucial role. Bacterial promoters, especially those of mesophilic bacteria, are in general preceded by a curved region. The marked thermosensitivity of curved DNA stretches allows bacteria to rapidly sense outer temperature variations and affects transcription by favoring the binding of activators or repressors. Curved DNA is also able to influence the transcriptional activity of a bacterial promoter directly, without the involvement of trans-acting regulators. This study attempts to quantitatively analyze the role of DNA curvature in thermoregulated gene expression using a real-time in vitro transcription model system based on a specific fluorescence molecular beacon. By analyzing the temperature-dependent expression of a reporter gene in a construct carrying a progressively decreasing bent sequence upstream from the promoter, we show that with a decrease in temperature a narrow curvature range accounts for a significant enhancement of promoter activity. This strengthens the view that DNA curvature-mediated regulation of gene expression is likely a strategy offering fine-tuning control possibilities and that, considering the widespread presence of curved sequences upstream from bacterial promoters, it may represent one of the most primitive forms of gene regulation.

摘要

在细菌迅速将其转录程序与新环境相匹配的分子策略中,序列介导的 DNA 弯曲变化起着至关重要的作用。细菌启动子,特别是那些嗜温细菌的启动子,通常前面有一个弯曲区域。弯曲 DNA 的显著热敏感性使细菌能够快速感知外部温度变化,并通过促进激活剂或抑制剂的结合来影响转录。弯曲 DNA 还能够直接影响细菌启动子的转录活性,而无需涉及反式作用调节剂。本研究试图使用基于特定荧光分子信标的实时体外转录模型系统,定量分析 DNA 弯曲在热调节基因表达中的作用。通过分析携带启动子上游逐渐减少弯曲序列的构建体中报告基因的温度依赖性表达,我们表明随着温度的降低,狭窄的曲率范围会显著增强启动子的活性。这加强了这样一种观点,即 DNA 弯曲介导的基因表达调控可能是一种提供微调控制可能性的策略,并且考虑到细菌启动子上游弯曲序列的广泛存在,它可能代表最原始的基因调控形式之一。

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