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-Structure paper
Title | Structural basis for target site selection in RNA-guided DNA transposition systems. |
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Journal, issue, pages | Science, Vol. 373, Issue 6556, Page 768-774, Year 2021 |
Publish date | Aug 13, 2021 |
Authors | Jung-Un Park / Amy Wei-Lun Tsai / Eshan Mehrotra / Michael T Petassi / Shan-Chi Hsieh / Ailong Ke / Joseph E Peters / Elizabeth H Kellogg / |
PubMed Abstract | 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. |
External links | Science / PubMed:34385391 / PubMed Central |
Methods | EM (helical sym.) / EM (single particle) |
Resolution | 3.2 - 4.2 Å |
Structure data | 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: |
Chemicals | ChemComp-AGS: ChemComp-MG: ChemComp-ADP: ChemComp-ATP: |
Source |
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Keywords | DNA BINDING PROTEIN/DNA / CRISPR / Transposition / AAA+ ATPase / Tn7 / DNA BINDING PROTEIN-DNA complex / DNA BINDING PROTEIN / Transposase |