Single-molecule dynamics of DNA gyrase in evolutionarily distant bacteria Mycobacterium tuberculosis and Escherichia coli.
Single-molecule dynamics of DNA gyrase in evolutionarily distant bacteria Mycobacterium tuberculosis and Escherichia coli.
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DOI：
10.1016/j.jbc.2023.103003
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发表时间：
2023-05
影响因子：
4.8
通讯作者：
Bryant, Zev
中科院分区：
文献类型：
--
作者：
Galvin, Cooper J.;Hobson, Matthew;Meng, Jonathan Xianglong;Ierokomos, Athena;Ivanov, Ivan E.;Berger, James M.;Bryant, Zev
DNA旋转酶是存在于所有细菌中的一种重要的核蛋白马达，也是结核分枝杆菌(MTB)感染抗生素治疗的主要靶点。用生物物理和生物化学方法研究过的链传递机制中，旋转酶可以将ATP水解酶的负超螺旋加到DNA上。为了分析导致链通过的子步骤的动力学，单分子转子磁珠跟踪(RBT)已经被用来跟踪大肠杆菌(EC)旋转酶的实时超螺旋和构象转变。然而，RBT还没有被应用于其他病原相关细菌的旋转酶，也不知道亚步是否在进化上遥远的物种中保守。在这里，我们比较了两个进化上遥远的细菌物种MTB和EC之间的旋转酶超螺旋动力学。我们使用RBT来测量纯化的旋转酶蛋白在与DNA的络合物中的超卷曲速率、过程以及构象状态的几何和转变动力学。结果表明，大肠埃希菌和结核分枝杆菌旋转酶都是进行性的，其中分枝杆菌酶的∼速度比EC酶慢5.5倍。与EC旋转酶相比，MTB旋转酶在存在和不存在ATP的情况下，也更容易形成DNA手性包裹在酶周围的中间状态。我们的子步测量揭示了EC和MTB旋转酶与DNA相互作用的构象状态的共同特征，但也表明了种群和过渡率的差异，这可能反映了这两个物种之间的不同细胞需求。
DNA gyrase is an essential nucleoprotein motor present in all bacteria and is a major target for antibiotic treatment of Mycobacterium tuberculosis (MTB) infection. Gyrase hydrolyzes ATP to add negative supercoils to DNA using a strand passage mechanism that has been investigated using biophysical and biochemical approaches. To analyze the dynamics of substeps leading to strand passage, single-molecule rotor bead tracking (RBT) has been used previously to follow real-time supercoiling and conformational transitions in Escherichia coli (EC) gyrase. However, RBT has not yet been applied to gyrase from other pathogenically relevant bacteria, and it is not known whether substeps are conserved across evolutionarily distant species. Here, we compare gyrase supercoiling dynamics between two evolutionarily distant bacterial species, MTB and EC. We used RBT to measure supercoiling rates, processivities, and the geometries and transition kinetics of conformational states of purified gyrase proteins in complex with DNA. Our results show that E. coli and MTB gyrases are both processive, with the MTB enzyme displaying velocities ∼5.5× slower than the EC enzyme. Compared with EC gyrase, MTB gyrase also more readily populates an intermediate state with DNA chirally wrapped around the enzyme, in both the presence and absence of ATP. Our substep measurements reveal common features in conformational states of EC and MTB gyrases interacting with DNA but also suggest differences in populations and transition rates that may reflect distinct cellular needs between these two species.
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影响因子：
16.8
作者：
Basu A;Schoeffler AJ;Berger JM;Bryant Z
通讯作者：
Bryant Z
影响因子：
14.9
作者：
Hobson, Matthew J.;Bryant, Zev;Berger, James M.
通讯作者：
Berger, James M.
影响因子：
14.9
作者：
Ashley RE;Dittmore A;McPherson SA;Turnbough CL Jr;Neuman KC;Osheroff N
通讯作者：
Osheroff N
DOI：
10.1073/pnas.93.25.14416
发表时间：
1996-12-10
影响因子：
11.1
作者：
Kampranis, SC;Maxwell, A
通讯作者：
Maxwell, A
DOI：
10.1016/j.bbrc.2006.07.017
发表时间：
2006-09-15
影响因子：
3.1
作者：
Aubry, Alexandra;Fisher, L. Mark;Cambau, Ernmanuelle
通讯作者：
Cambau, Ernmanuelle