8BD4
TniQ-capped Tns-ATP-dsDNA complex
Summary for 8BD4
| Entry DOI | 10.2210/pdb8bd4/pdb |
| EMDB information | 15974 |
| Descriptor | TnsC, TniQ (Homology model), DNA (5'-D(P*GP*AP*TP*CP*GP*AP*TP*CP*GP*AP*TP*CP*GP*AP*TP*C)-3'), ... (6 entities in total) |
| Functional Keywords | transposition, tniq, tnsc, crispr-cas, tn7-like transposon, dna binding protein |
| Biological source | Scytonema hofmannii More |
| Total number of polymer chains | 12 |
| Total formula weight | 291055.28 |
| Authors | Querques, I.,Schmitz, M.,Oberli, S.,Chanez, C.,Jinek, M. (deposition date: 2022-10-18, release date: 2022-12-28, Last modification date: 2024-07-24) |
| Primary citation | Schmitz, M.,Querques, I.,Oberli, S.,Chanez, C.,Jinek, M. Structural basis for the assembly of the type V CRISPR-associated transposon complex. Cell, 185:4999-, 2022 Cited by PubMed Abstract: 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. PubMed: 36435179DOI: 10.1016/j.cell.2022.11.009 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.44 Å) |
Structure validation
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