A New Member of MocR/GabR-type PLP-Binding Regulator of D-Alanyl-D-Alanine Ligase in Brevibacillus brevis.
A New Member of MocR/GabR-type PLP-Binding Regulator of D-Alanyl-D-Alanine Ligase in Brevibacillus brevis.
复制标题
短芽孢杆菌 D-丙氨酰-D-丙氨酸连接酶 MocR/GabR 型 PLP 结合调节因子的新成员。
DOI：
10.1111/febs.13415
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Yoshimura T.
中科院分区：
文献类型：
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作者：
Takenaka T.;Ito T.;Miyahara I.;Hemmi H.;Yoshimura T.
短芽孢杆菌 BBR47_28440 基因（称为 asddlR）编码 MocR/GabR 家族转录调节因子，由 N 端螺旋-转角-螺旋 DNA 结合结构域和 C 端转氨酶样结构域组成。 TheddlR 基因位于 theB 中 d-丙氨酰-d-丙氨酸连接酶基因 (ddl) 的上游。 brevisgenome，这两个基因形成操纵子。凝胶迁移测定表明，纯化的 DdlR 特异性结合 DNA 区域，其中包括 ddlR 启动子的推定的 -35 和 -10 区域。发现与 theddlR 启动子 -10 区域重叠的 6 bp 反向重复序列对于结合很重要。体内报告基因检测证实 DdlR 是 theddlR-ddloperon 的激活剂。光谱分析表明，纯化的 DdlR 是一种吡哆醛 5'-磷酸结合转录调节因子，具有 Ddl 的酶促产物福特-丙氨酰-d-丙氨酸和甘氨酰甘氨酸的二肽结合能力。 DdlR 能够形成二肽-吡哆醛 5'-磷酸外醛亚胺，但缺乏转氨酶活性。生物信息学分析表明，DdlR 介导的 ddlRandddl 转录调控可能发生在多种细菌系统中，例如放线菌和芽孢杆菌属。
TheBrevibacillus brevis BBR47_28440gene (referred to asddlR) encodes an MocR/GabR family transcriptional regulator consisting of an N‐terminal helix‐turn‐helix DNA binding domain and a C‐terminal aminotransferase‐like domain. TheddlRgene is located just upstream of thed‐alanyl‐d‐alanine ligase gene (ddl) in theB. brevisgenome, and these two genes form an operon. Gel‐shift assays indicated that purified DdlR binds specifically to the DNA region that includes putative −35 and −10 regions of theddlRpromoter. A 6‐bp inverted repeat that overlaps the −10 region of theddlRpromoter was found to be important for the binding.In vivoreporter assays confirmed that DdlR is an activator of theddlR‐ddloperon. Spectroscopic analyses indicated that purified DdlR is a pyridoxal 5′‐phosphate binding transcriptional regulator that has dipeptide binding ability ford‐alanyl‐d‐alanine, the enzymatic product of Ddl, and glycylglycine. DdlR is capable of forming a dipeptide‐pyridoxal 5′‐phosphate external aldimine, but it lacks aminotransferase activity. Bioinformatic analyses suggest that DdlR‐mediated transcriptional regulation ofddlRandddlmay occur in multiple bacterial systems such as Actinobacteria andBacillusspecies.