Cueny Rachel R, Voter Andrew F, McKenzie Aidan M, Morgenstern Marcel, Myers Kevin S, Place Michael M, Peters Jason M, Coon Joshua J, Keck James L
Biomolecular Chemistry Department, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI, 53706 United States.
National Center for Quantitative Biology of Complex Systems, University of Wisconsin-Madison, Madison, WI, 53706 United States.
Nucleic Acids Res. 2025 Mar 20;53(6). doi: 10.1093/nar/gkaf264.
G-quadruplex (G4) structures can form in guanine-rich DNA or RNA and have been found to modulate cellular processes, including replication, transcription, and translation. Many studies on the cellular roles of G4s have focused on eukaryotic systems, with far fewer probing bacterial G4s. Using a chemical-genetic approach, we identified genes in Escherichia coli that are important for growth in G4-stabilizing conditions. Reducing levels of translation elongation factor Tu or slowing translation initiation or elongation with kasugamycin, chloramphenicol, or spectinomycin suppress the effects of G4-stabilizing compounds. In contrast, reducing the expression of specific translation termination or ribosome recycling proteins is detrimental to growth in G4-stabilizing conditions. Proteomic and transcriptomic analyses reveal decreased protein and transcript levels, respectively, for ribosome assembly factors and proteins associated with translation in the presence of G4 stabilizer. Our results support a model in which reducing the rate of translation by altering translation initiation, translation elongation, or ribosome assembly can compensate for G4-related stress in E. coli.
G-四链体（G4）结构可在富含鸟嘌呤的DNA或RNA中形成，并且已发现其可调节细胞过程，包括复制、转录和翻译。许多关于G4细胞作用的研究都集中在真核系统上，而对细菌G4的探究要少得多。我们采用化学遗传学方法，在大肠杆菌中鉴定出了对在G4稳定条件下生长至关重要的基因。降低翻译延伸因子Tu的水平，或用春日霉素、氯霉素或壮观霉素减缓翻译起始或延伸，可抑制G4稳定化合物的作用。相比之下，降低特定翻译终止或核糖体循环蛋白的表达对在G4稳定条件下的生长有害。蛋白质组学和转录组学分析分别揭示，在存在G4稳定剂的情况下，核糖体组装因子和与翻译相关的蛋白质的蛋白质水平和转录水平均降低。我们的结果支持这样一种模型，即通过改变翻译起始、翻译延伸或核糖体组装来降低翻译速率，可以补偿大肠杆菌中与G4相关的应激。
