Characterization of the MurT/GatD complex in Mycobacterium tuberculosis towards validating a novel anti-tubercular drug target.
Characterization of the MurT/GatD complex in Mycobacterium tuberculosis towards validating a novel anti-tubercular drug target.
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结核分枝杆菌中MurT/ gad复合体的特性对新型抗结核药物靶点的验证
DOI：
10.1093/jacamr/dlab028
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发表时间：
2021-03
影响因子：
3.4
通讯作者：
Bhakta S
中科院分区：
文献类型：
--
作者：
Maitra A;Nukala S;Dickman R;Martin LT;Munshi T;Gupta A;Shepherd AJ;Arnvig KB;Tabor AB;Keep NH;Bhakta S
识别和验证新的治疗靶点对于解决结核病耐药性的上升至关重要。一个重要的Mur连接酶样基因（Rv 3712），预计将参与细胞壁肽聚糖（PG）的生物合成和保守的分枝杆菌，包括基因耗尽麻风分枝杆菌，是本研究的主要重点。使用无机磷酸盐释放测定法进行Rv 3712的生化分析。操纵子的结构进行了鉴定，使用逆转录酶PCR和转录/翻译融合载体。体内分枝杆菌蛋白质片段互补测定有助于产生相互作用组。Rv 3712是一种ATP酶。其操纵子的表征揭示了驱动Rv 3712和Rv 3713共转录的分枝杆菌特异性启动子。发现这两种基因产物在体内相互作用。基于序列的功能分配显示Rv 3712和Rv 3713可能分别是分枝杆菌PG受体修饰酶MurT和GatD。还鉴定了涉及Mtb-MurT、调节蛋白和细胞分裂蛋白的体内网络。了解酶复合物在PG代谢和细胞分裂中的作用，以及对抗菌素耐药性和宿主免疫反应的影响，将有助于设计特异性针对M的治疗药物。结核
Identification and validation of novel therapeutic targets is imperative to tackle the rise of drug resistance in tuberculosis. An essential Mur ligase-like gene (Rv3712), expected to be involved in cell-wall peptidoglycan (PG) biogenesis and conserved across mycobacteria, including the genetically depleted Mycobacterium leprae, was the primary focus of this study. Biochemical analysis of Rv3712 was performed using inorganic phosphate release assays. The operon structure was identified using reverse-transcriptase PCR and a transcription/translation fusion vector. In vivo mycobacterial protein fragment complementation assays helped generate the interactome. Rv3712 was found to be an ATPase. Characterization of its operon revealed a mycobacteria-specific promoter driving the co-transcription of Rv3712 and Rv3713. The two gene products were found to interact with each other in vivo. Sequence-based functional assignments reveal that Rv3712 and Rv3713 are likely to be the mycobacterial PG precursor-modifying enzymes MurT and GatD, respectively. An in vivo network involving Mtb-MurT, regulatory proteins and cell division proteins was also identified. Understanding the role of the enzyme complex in the context of PG metabolism and cell division, and the implications for antimicrobial resistance and host immune responses will facilitate the design of therapeutics that are targeted specifically to M. tuberculosis.