Structural basis of transposon end recognition explains central features of Tn7 transposition systems.

Structural basis of transposon end recognition explains central features of Tn7 transposition systems.
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DOI:
10.1016/j.molcel.2022.05.005
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发表时间:
2022-07-21
期刊:
影响因子:
16
通讯作者:
Nowotny, Marcin
Nowotny, Marcin
中科院分区:
生物学1区
文献类型:
--
作者:
Kaczmarska, Zuzanna;Czarnocki-Cieciura, Mariusz;Gorecka-Minakowska, Karolina M.;Wingo, Robert J.;Jackiewicz, Justyna;Zajko, Weronika;Poznanski, Jaroslaw T.;Rawski, Michal;Grant, Timothy;Peters, Joseph E.;Nowotny, Marcin

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Tn 7是一种细菌转座子,其亲属含有介导RNA引导的转座子插入的元件编码的CRISPR-Cas系统。在这里,我们提出的2.7倍低温电子显微镜结构的原型Tn 7转座酶TnsB与转座子末端DNA相互作用。当TnsB跨重复结合位点相互作用时,它采用串珠结构,其中DNA结合和催化结构域以平铺和交织的方式排列。DNA结合结构域形成很少的碱基特异性接触,导致结合偏好,需要多个弱保守位点在适当的间距,以实现DNA序列特异性。TnsB结合赋予由结合位点的间隔或重叠决定的蛋白质结合的DNA末端的全局结构的差异,解释元件的左端和右端的功能差异。我们提出了一个模型的链转移复合物,其中的终端TnsB分子重排,使其催化结构域是在一个有利于转座的位置。Kaczmarska等人提出了与转座子末端DNA结合的Tn 7转座子转座酶TnsB的结构。Tn 7是一种细菌转座子。其含有CRISPR-Cas机制的亲戚是基因编辑的有前途的工具。TnsB分子与转座子末端中的重复结合位点以交织的方式相互作用以进行特异性末端识别。
Tn7 is a bacterial transposon with relatives containing element-encoded CRISPR-Cas systems mediating RNA-guided transposon insertion. Here, we present the 2.7 Å cryoelectron microscopy structure of prototypic Tn7 transposase TnsB interacting with the transposon end DNA. When TnsB interacts across repeating binding sites, it adopts a beads-on-a-string architecture, where the DNA-binding and catalytic domains are arranged in a tiled and intertwined fashion. The DNA-binding domains form few base-specific contacts leading to a binding preference that requires multiple weakly conserved sites at the appropriate spacing to achieve DNA sequence specificity. TnsB binding imparts differences in the global structure of the protein-bound DNA ends dictated by the spacing or overlap of binding sites explaining functional differences in the left and right ends of the element. We propose a model of the strand-transfer complex in which the terminal TnsB molecule is rearranged so that its catalytic domain is in a position conducive to transposition. Kaczmarska et al. present a structure of Tn7 transposon transposase TnsB bound to transposon end DNA. Tn7 is a bacterial transposon. Its relatives containing CRISPR-Cas machinery are promising tools for gene editing. TnsB molecules interact with repeating binding sites in the transposon end in an intertwined fashion for specific end recognition.
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