Structural basis for target site selection in RNA-guided DNA transposition systems

doi:10.1126/science.abi8976

GSTDTAP > 气候变化

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	被引频次：35[WOS] [WOS记录] [WOS相关记录]
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).