Establishment of titration-based control of DNA replication in Escherichia coli

IF 13.9 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Science Advances Pub Date : 2026-10-16 DOI:10.1126/sciadv.aei9788

Belén Adiego-Pérez, Christina Ludwig, Demian Fluit, Mareike Berger, Alejandra Alonso Santirso, Johannes Hohlbein, Raymond H. J. Staals, Pieter Rein ten Wolde, John van der Oost, Nico J. Claassens, Lorenzo Olivi

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Abstract

Escherichia coli couples the initiation of DNA replication with cell size by modulating the activity of the replication initiator protein DnaA. The activity of DnaA is regulated both by its interconversion between an active and inactive form and by its titration on chromosomal binding sites. Whereas its interconversion has been thoroughly studied, the extent to which DnaA titration can control replication initiation is poorly understood. Here, we describe the control of E. coli DNA replication via titration by modulating the expression of a constitutively active DnaA variant in four growth conditions. While we obtained stable cell cycles during slow growth, faster growth associated with overlapping replication forks led to initiation instability and replicative stress. Overall, our results provide insights into the limits of titration-based systems in the control of genome replication and their potential role in the evolutionary trajectory of E. coli. Last, this study provides design principles for a simplified, titration-only regulatory mechanism for DNA replication in synthetic cells.

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建立基于滴定法的大肠杆菌DNA复制控制方法

大肠杆菌通过调节复制起始蛋白DNA的活性，将DNA复制的起始与细胞大小偶联。dna的活性是通过其在活性形式和非活性形式之间的相互转化以及在染色体结合位点上的滴定来调节的。而其互变现象已彻底研究,DnaA滴定的程度可以控制复制起始知之甚少。在这里，我们描述了控制大肠杆菌DNA复制通过滴定通过调节组成活性DNA变体的表达在四种生长条件。虽然我们在缓慢生长过程中获得了稳定的细胞周期，但与重叠复制分叉相关的快速生长导致起始不稳定和复制应激。总的来说，我们的结果提供了基于滴定的系统在控制基因组复制方面的局限性及其在大肠杆菌进化轨迹中的潜在作用的见解。最后，本研究为合成细胞中DNA复制的简化、仅滴定调节机制提供了设计原则。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Science Advances 综合性期刊-综合性期刊

CiteScore

21.40

自引率

1.50%

发文量

1937

审稿时长

29 weeks

期刊介绍： Science Advances, an open-access journal by AAAS, publishes impactful research in diverse scientific areas. It aims for fair, fast, and expert peer review, providing freely accessible research to readers. Led by distinguished scientists, the journal supports AAAS's mission by extending Science magazine's capacity to identify and promote significant advances. Evolving digital publishing technologies play a crucial role in advancing AAAS's global mission for science communication and benefitting humankind.