Distinct Mechanism Evolved for Mycobacterial RNA Polymerase and Topoisomerase I Protein-Protein Interaction.
Distinct Mechanism Evolved for Mycobacterial RNA Polymerase and Topoisomerase I Protein-Protein Interaction.
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分枝杆菌 RNA 聚合酶和拓扑异构酶 I 蛋白质-蛋白质相互作用的独特机制已进化。
DOI：
10.1016/j.jmb.2017.08.011
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发表时间：
2017-09-15
影响因子：
5.6
通讯作者：
Tse-Dinh YC
中科院分区：
文献类型：
--
作者：
Banda S;Cao N;Tse-Dinh YC
我们在这里报道了在结核分枝杆菌和污垢分枝杆菌中拓扑异构酶I和RNA聚合酶之间相互作用的独特机制，它是独立于先前描述的细菌拓扑异构酶I和RNA聚合酶之间的相互作用而进化的。细菌DNA拓扑异构酶I负责防止基因组DNA的超负超螺旋。在转录延伸过程中，拓扑异构酶I与RNA聚合酶的结合可以有效地缓解转录驱动的负超卷曲。我们的结果表明，在没有β的情况下，拓扑异构酶I的C末端结构域(TOPOI-CTD)与耻垢分枝杆菌核糖核酸聚合酶的DNA亚单位之间存在直接的物理作用。在进化上，分枝杆菌中的Topoi-CTD在氨基酸序列和三维结构上与大多数放线菌外的细菌物种中发现的Topoi-CTD无关，包括大肠杆菌。在分枝杆菌和大肠杆菌中，拓扑异构酶I和RNA聚合酶之间的功能相互作用是独立进化的，这种蛋白质-蛋白质相互作用所利用的拓扑异构酶I的结构元件截然不同。大肠杆菌TOPOI-CTD中的锌带基序参与了与RNA聚合酶的相互作用。对于耻垢分枝杆菌Topoi-CTD，C末端富含碱性残基的27个氨基酸的尾巴负责与RNA聚合酶的相互作用。在耻垢分枝杆菌中过表达的Topoi-CTD与内源拓扑异构酶I竞争蛋白质与RNA聚合酶的相互作用。Topoi-CTD的过表达导致抗生素和过氧化氢处理后存活率下降，支持了拓扑异构酶I和RNA聚合酶之间的蛋白质-蛋白质相互作用在分枝杆菌应激反应中的重要性。
We report here a distinct mechanism of interaction between topoisomerase I and RNA polymerase in Mycobacterium tuberculosis and Mycobacterium smegmatis that has evolved independently from the previously characterized interaction between bacterial topoisomerase I and RNA polymerase. Bacterial DNA topoisomerase I is responsible for preventing the hyper-negative supercoiling of genomic DNA. The association of topoisomerase I with RNA polymerase during transcription elongation could efficiently relieve transcription-driven negative supercoiling. Our results demonstrate a direct physical interaction between the C-terminal domains of topoisomerase I (TopoI-CTD) and the β′ subunit of RNA polymerase of M. smegmatis in the absence of DNA. The TopoI-CTD in mycobacteria are evolutionarily unrelated in amino acid sequence and three-dimensional structure to the TopoI-CTD found in the majority of bacterial species outside Actinobacteria, including Escherichia coli. The functional interaction between topoisomerase I and RNA polymerase has evolved independently in mycobacteria and E. coli, with distinctively different structural elements of TopoI-CTD utilized for this protein-protein interaction. Zinc ribbon motifs in E. coli TopoI-CTD are involved in the interaction with RNA polymerase. For M. smegmatis TopoI-CTD, a 27 amino acid tail that is rich in basic residues at the C-terminal end is responsible for the interaction with RNA polymerase. Overexpression of recombinant TopoI-CTD in M. smegmatis competed with the endogenous topoisomerase I for protein-protein interactions with RNA polymerase. The TopoI-CTD overexpression resulted in decreased survival following treatment with antibiotics and hydrogen peroxide, supporting the importance of the protein-protein interaction between topoisomerase I and RNA polymerase during stress response of mycobacteria.
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