Akiyama Tomonori, Yamada Yusuke, Takaya Naoki, Ito Shinsaku, Sasaki Yasuyuki, Yajima Shunsuke
Department of Bioscience, Tokyo University of Agriculture, Setagaya-ku, Tokyo 156-8502, Japan.
Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization, 1-1 Oho, Tsukuba 305-0801, Japan.
Acta Crystallogr F Struct Biol Commun. 2017 Jan 1;73(Pt 1):16-23. doi: 10.1107/S2053230X16019208.
The bacterial transcription factor IclR (isocitrate lyase regulator) is a member of a one-component signal transduction system, which shares the common motif of a helix-turn-helix (HTH)-type DNA-binding domain (DBD) connected to a substrate-binding domain (SBD). Here, the crystal structure of an IclR homologue (Mi-IclR) from Microbacterium sp. strain HM58-2, which catabolizes acylhydrazide as the sole carbon source, is reported. Mi-IclR is expected to regulate an operon responsible for acylhydrazide degradation as an initial step. Native single-wavelength anomalous diffraction (SAD) experiments were performed in combination with molecular replacement. CRANK2 from the CCP4 suite successfully phased and modelled the complete structure of a homotetramer composed of 1000 residues in an asymmetric unit, and the model was refined to 2.1 Å resolution. The overall structure of Mi-IclR shared the same domain combination as other known IclR structures, but the relative geometry between the DBD and SBD differs. Accordingly, the geometry of the Mi-IclR tetramer was unique: the putative substrate-binding site in each subunit is accessible from the outside of the tetramer, as opposed to buried inside as in the previously known IclR structures. These differences in the domain geometry may contribute to the transcriptional regulation of IclRs.
细菌转录因子IclR(异柠檬酸裂合酶调节因子)是单组分信号转导系统的成员,该系统具有与底物结合结构域(SBD)相连的螺旋-转角-螺旋(HTH)型DNA结合结构域(DBD)的共同基序。在此,报道了来自微杆菌属菌株HM58-2的IclR同源物(Mi-IclR)的晶体结构,该菌株以酰肼作为唯一碳源进行分解代谢。预计Mi-IclR作为第一步调节负责酰肼降解的操纵子。结合分子置换进行了天然单波长反常衍射(SAD)实验。CCP4套件中的CRANK2成功解析并构建了一个不对称单元中由1000个残基组成的同四聚体的完整结构模型,并将该模型精修至2.1 Å分辨率。Mi-IclR的整体结构与其他已知的IclR结构具有相同的结构域组合,但DBD和SBD之间的相对几何形状不同。因此,Mi-IclR四聚体的几何形状是独特的:与先前已知的IclR结构中埋在内部不同,每个亚基中假定的底物结合位点可从四聚体外部进入。这些结构域几何形状的差异可能有助于IclR的转录调控。