8bd5

Cas12k-sgRNA-dsDNA-S15-TniQ-TnsC transposon recruitment complex

spinqualitylabelsall models 8bd5, resolution 3.30Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 8bd5 is a 13 chain structure with sequence from Escherichia coli K-12, Scytonema hofmannii and Synthetic construct. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: Electron Microscopy, Resolution 3.3Å Ligands: ATP, MG, ZN Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function A0A8X6EH11_9CYAN Publication Abstract from PubMed CRISPR-Cas systems have been co-opted by Tn7-like transposable 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 comprised of RNA-guided Cas12k, TniQ, a polymeric TnsC filament and, unexpectedly, the ribosomal protein S15. Complex assembly, mediated by a network of interactions involving the guide RNA, TniQ, and S15, results in R-loop completion. TniQ contacts two TnsC protomers at the Cas12k-proximal filament end, likely nucleating its polymerization. Transposition activity assays corroborate our structural findings, implying that S15 is a bona fide component of the type V crRNA-guided transposon machinery. Altogether, our work uncovers key mechanistic aspects underpinning RNA-mediated assembly of CRISPR-associated transposons to guide their development as programmable tools for site-specific insertion of large DNA payloads. Structural basis for the assembly of the type V CRISPR-associated transposon complex.,Schmitz M, Querques I, Oberli S, Chanez C, Jinek M Cell. 2022 Dec 22;185(26):4999-5010.e17. doi: 10.1016/j.cell.2022.11.009. Epub , 2022 Nov 25. PMID:36435179 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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