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-Structure paper
タイトル | Structural basis for target site selection in RNA-guided DNA transposition systems. |
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ジャーナル・号・ページ | Science, Vol. 373, Issue 6556, Page 768-774, Year 2021 |
掲載日 | 2021年8月13日 |
著者 | Jung-Un Park / Amy Wei-Lun Tsai / Eshan Mehrotra / Michael T Petassi / Shan-Chi Hsieh / Ailong Ke / Joseph E Peters / Elizabeth H Kellogg / |
PubMed 要旨 | 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 ...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•AlF These mechanistic findings provide the underpinnings for engineering CRISPR-associated transposition systems for research and therapeutic applications. |
リンク | Science / PubMed:34385391 / PubMed Central |
手法 | EM (らせん対称) / EM (単粒子) |
解像度 | 3.2 - 4.2 Å |
構造データ | EMDB-23720: ATPgS bound TnsC filament from shCAST system EMDB-23721, PDB-7m9a: EMDB-23722, PDB-7m9b: EMDB-23723, PDB-7m9c: EMDB-23724: EMDB-23725: EMDB-23726, PDB-7n6i: |
化合物 | ChemComp-AGS: ChemComp-MG: ChemComp-ADP: ChemComp-ATP: |
由来 |
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キーワード | DNA BINDING PROTEIN/DNA / CRISPR / Transposition / AAA+ ATPase / Tn7 / DNA BINDING PROTEIN-DNA complex / DNA BINDING PROTEIN / Transposase |