Institute of Bioscience and Biotechnology, Kangwon National University, 1 Kangwondaehakgil, Chuncheon 24341, Republic of Korea.
Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, 1 Kangwondaehakgil, Chuncheon 24341, Republic of Korea.
Nucleic Acids Res. 2024 Nov 27;52(21):13192-13205. doi: 10.1093/nar/gkae922.
Uric acid (UA)-responsive transcriptional regulators (UrtRs), which belong to the multiple antibiotic resistance regulator (MarR) superfamily, transcriptionally coordinate virulence and metabolism in bacteria by modulating interactions with operator DNA in response to UA. To elucidate the transcriptional regulatory mechanism of UrtR, we structurally analyzed UrtR proteins, including PecS, MftR, and HucR, alone and in complex with UA or DNA. UrtR contains a dimerization domain (DD) and a winged helix-turn-helix domain (wHTHD) and forms a homodimer primarily via the DD, as observed for other MarR superfamily proteins. However, UrtRs are characterized by a unique N-terminal α-helix, which contributes to dimerization and UA recognition. In the absence of UA, the UrtR dimer symmetrically binds to the operator double-stranded DNA (dsDNA) by inserting its α4 recognition helix and β-stranded wing within the wHTHD into the major and minor grooves of dsDNA, respectively. Upon exposure to UA, UrtR accommodates UA in the intersubunit pocket between the DD and wHTHD. UA binding induces a conformational change in the major groove-binding core element of the UrtR wHTHD, generating a DNA binding-incompatible structure. This local allosteric mechanism of UrtR completely differs from that generally observed in other MarR superfamily members, in which the entire wHTHD undergoes effector-responsive global shifts.
尿酸(UA)反应性转录调节因子(UrtR)属于多重抗生素耐药调节因子(MarR)超家族,通过调节与 UA 反应的操纵子 DNA 的相互作用,转录协调细菌的毒力和代谢。为了阐明 UrtR 的转录调控机制,我们对 PecS、MftR 和 HucR 等 UrtR 蛋白进行了结构分析,单独和与 UA 或 DNA 复合。UrtR 包含一个二聚化结构域(DD)和一个翼型螺旋-转角-螺旋结构域(wHTHD),主要通过 DD 形成同源二聚体,就像其他 MarR 超家族蛋白一样。然而,UrtR 的特征是具有独特的 N 端α-螺旋,有助于二聚体形成和 UA 识别。在没有 UA 的情况下,UrtR 二聚体通过将其α4 识别螺旋和β链状翼插入 wHTHD 中的大、小沟,对称地结合到操纵子双链 DNA(dsDNA)上,分别。UA 暴露后,UrtR 将 UA 容纳在 DD 和 wHTHD 之间的亚基间口袋中。UA 结合诱导 UrtR wHTHD 的大沟结合核心元件发生构象变化,产生与 DNA 结合不兼容的结构。这种 UrtR 的局部变构机制与其他 MarR 超家族成员中通常观察到的机制完全不同,在其他 MarR 超家族成员中,整个 wHTHD 会发生效应物响应的全局位移。