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Title: | Structural basis for RNA-mediated assembly of type V CRISPR-associated transposons |
Authors: | Schmitz, Michael |
Keywords: | Cơ sở cấu trúc; lắp ráp; trung gian RNA; transposon; CRISPR loại V |
Issue Date: | 2023 |
Publisher: | bioRxiv |
Abstract: | CRISPR systems have been co-opted by Tn7-like elements to direct RNA-guided transposition. Type V-K CRISPR-associated transposons rely on the concerted activities of the pseudonuclease Cas12k, the AAA+ ATPase TnsC, the Zn-finger protein TniQ, and the transposase TnsB. Here we present a cryo-electron microscopic structure of a target DNA-bound Cas12k-transposon recruitment complex comprising RNA-guided Cas12k, TniQ, TnsC and, unexpectedly, the ribosomal protein S15. Complex assembly on target DNA results in complete R-loop formation mediated by critical interactions between TniQ and the trans-activating crRNA, and is coupled with TniQ-dependent nucleation of a TnsC filament. In vivo transposition assays corroborate our structural findings, and biochemical and functional analyses of S15 supports its role as a bona fide component of the type V crRNA-guided transposition machinery. |
URI: | http://dlib.hust.edu.vn/handle/HUST/23552 |
Link item primary: | https://www.biorxiv.org/content/10.1101/2022.06.17.496590v1.full.pdf+html |
Appears in Collections: | OER - Kỹ thuật hóa học; Công nghệ sinh học - Thực phẩm; Công nghệ môi trường |
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