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DOI | 10.1126/science.abi8976 |
Structural basis for target site selection in RNA-guided DNA transposition systems | |
Jung-Un Park; Amy Wei-Lun Tsai; Eshan Mehrotra; Michael T. Petassi; Shan-Chi Hsieh; Ailong Ke; Joseph E. Peters; Elizabeth H. Kellogg | |
2021-08-13 | |
发表期刊 | Science |
出版年 | 2021 |
英文摘要 | Exciting genomic engineering possibilities exist for natural integration systems called transposons, which have coopted CRISPR/Cas systems. An unexplained feature of these systems involves how they direct insertions in a single orientation at a precise distance from the programmed target sequence. Park et al. show that orientation information is communicated to the transposase TnsB using the unidirectional growth of a helical filament made up of an AAA+ protein, TnsC. ATP hydrolysis trims the filament to a minimal unit that is marked by TniQ and defined by the Cas12k protein to provide spacing information. This finding may help future engineering of these systems for therapeutic applications. Science , abi8976, this issue p. [768][1] CRISPR-associated transposition systems allow guide RNA–directed integration of a single DNA cargo in one orientation at a fixed distance from a programmable target sequence. We used cryo–electron microscopy (cryo-EM) to define the mechanism that underlies this process by characterizing the transposition regulator, TnsC, from a type V-K CRISPR-transposase system. In this scenario, polymerization of adenosine triphosphate–bound TnsC helical filaments could explain how polarity information is passed to the transposase. TniQ caps the TnsC filament, representing a universal mechanism for target information transfer in Tn7/Tn7-like elements. Transposase-driven disassembly establishes delivery of the element only to unused protospacers. Finally, TnsC transitions to define the fixed point of insertion, as revealed by structures with the transition state mimic ADP•AlF3. These mechanistic findings provide the underpinnings for engineering CRISPR-associated transposition systems for research and therapeutic applications. [1]: /lookup/doi/10.1126/science.abi8976 |
领域 | 气候变化 ; 资源环境 |
URL | 查看原文 |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://119.78.100.173/C666/handle/2XK7JSWQ/335879 |
专题 | 气候变化 资源环境科学 |
推荐引用方式 GB/T 7714 | Jung-Un Park,Amy Wei-Lun Tsai,Eshan Mehrotra,et al. Structural basis for target site selection in RNA-guided DNA transposition systems[J]. Science,2021. |
APA | Jung-Un Park.,Amy Wei-Lun Tsai.,Eshan Mehrotra.,Michael T. Petassi.,Shan-Chi Hsieh.,...&Elizabeth H. Kellogg.(2021).Structural basis for target site selection in RNA-guided DNA transposition systems.Science. |
MLA | Jung-Un Park,et al."Structural basis for target site selection in RNA-guided DNA transposition systems".Science (2021). |
条目包含的文件 | 条目无相关文件。 |
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