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-Structure paper
タイトル | Conformational landscape of the type V-K CRISPR-associated transposon integration assembly. |
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ジャーナル・号・ページ | Mol Cell, Vol. 84, Issue 12, Page 2353-22367.e5, Year 2024 |
掲載日 | 2024年6月20日 |
著者 | Francisco Tenjo-Castaño / Nicholas Sofos / Luisa S Stutzke / Piero Temperini / Anders Fuglsang / Tillmann Pape / Pablo Mesa / Guillermo Montoya / |
PubMed 要旨 | CRISPR-associated transposons (CASTs) are mobile genetic elements that co-opt CRISPR-Cas systems for RNA-guided DNA transposition. CASTs integrate large DNA cargos into the attachment (att) site ...CRISPR-associated transposons (CASTs) are mobile genetic elements that co-opt CRISPR-Cas systems for RNA-guided DNA transposition. CASTs integrate large DNA cargos into the attachment (att) site independently of homology-directed repair and thus hold promise for eukaryotic genome engineering. However, the functional diversity and complexity of CASTs hinder an understanding of their mechanisms. Here, we present the high-resolution cryoelectron microscopy (cryo-EM) structure of the reconstituted ∼1 MDa post-transposition complex of the type V-K CAST, together with different assembly intermediates and diverse TnsC filament lengths, thus enabling the recapitulation of the integration complex formation. The results of mutagenesis experiments probing the roles of specific residues and TnsB-binding sites show that transposition activity can be enhanced and suggest that the distance between the PAM and att sites is determined by the lengths of the TnsB C terminus and the TnsC filament. This singular model of RNA-guided transposition provides a foundation for repurposing the system for genome-editing applications. |
リンク | Mol Cell / PubMed:38834066 |
手法 | EM (単粒子) |
解像度 | 2.3 - 3.19 Å |
構造データ | EMDB-15697, PDB-8axa: EMDB-19075, PDB-8rdu: EMDB-19282, PDB-8rkt: EMDB-19283, PDB-8rku: EMDB-19284, PDB-8rkv: EMDB-19286: consensus map of the V-K CRISPR-associated Transposon Integration Assembly |
化合物 | ChemComp-HOH: ChemComp-MG: ChemComp-ZN: ChemComp-ATP: |
由来 |
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キーワード | DNA BINDING PROTEIN / CRISPR / complex / transposition / CRISPR-associated Transposon genome editing transposition |