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首页> 外文期刊>Journal of Molecular Biology >Distinct Mechanism Evolved for Mycobacterial RNA Polymerase and Topoisomerase I Protein-Protein Interaction
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Distinct Mechanism Evolved for Mycobacterial RNA Polymerase and Topoisomerase I Protein-Protein Interaction

机译：用于分枝杆菌RNA聚合酶和拓扑异构酶I蛋白质 - 蛋白质相互作用的明显机制

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摘要

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 (Topol-CTDs) and the f3' subunit of RNA polymerase of M. smegmatis in the absence of DNA. The Topol-CTDs in mycobacteria are evolutionarily unrelated in amino acid sequence and three-dimensional structure to the Topol-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 Topol-CTD utilized for this protein protein interaction. Zinc ribbon motifs in E. coliTopol-CTD are involved in the interaction with RNA polymerase. For M. smegmatis Topol-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 Topol-CTD in M. smegmatis competed with the endogenous topoisomerase I for protein protein interactions with RNA polymerase. The Topol-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. (C) 2017 Elsevier Ltd. All rights reserved.
机译：在这里，在此报告拓扑异构酶I和RNA聚合酶在结核分枝杆菌和分枝杆菌聚合酶之间的不同机制，该分枝杆菌和分枝杆菌的分枝杆菌与细菌拓扑酶I和RNA聚合酶之间的先前表征相互作用不同。细菌DNA拓扑异构酶I负责预防基因组DNA的超阴性超晶体。在转录伸长期间与RNA聚合酶具有RNA聚合酶的衔接子酶的缔合可以有效地缓解转录驱动的阴性超铜。我们的结果表明，在不存在DNA的情况下，甲基菌酶I（TOPOL-CTDS）和RNA聚合酶的F3'亚基之间的直接物理相互作用。分枝杆菌中的蜂鸣器CTD在氨基酸序列和三维结构中进化地与猕猴菌外部大多数细菌种类中发现的蜂鸣CTD无关，包括大肠杆菌。拓扑异构酶I和RNA聚合酶之间的功能相互作用在分枝杆菌和大肠杆菌中独立地演化，具有用于该蛋白质蛋白质相互作用的蜂鸣器-CTD的独特不同的结构元素。 E.Colitopol-CTD中的锌带基序参与与RNA聚合酶的相互作用。对于M. Smogmatis Topol-CTD，在C末端富含碱性残留物的27-氨基酸尾部负责与RNA聚合酶的相互作用。在M. Smogmatis中重组蜂鸣体CTD的过度表达与内源性拓扑异构酶I竞争，用于与RNA聚合酶的蛋白质蛋白质相互作用。在用抗生素和过氧化氢处理后，蜂鸣器-CTD过表达导致存活率降低，支持在分枝杆菌的应激响应期间拓扑异构酶I和RNA聚合酶之间的蛋白质蛋白质相互作用的重要性。 （c）2017 Elsevier Ltd.保留所有权利。

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