Unveiling Cas8 dynamics and regulation in a transposon-encoded Cascade-TniQ Complex

We present detailed structural models of the RNA- and DNA-bound Cascade-TniQ complex, as described by Patel et al. in “Unveiling Cas8 Dynamics and Regulation in a Transposon-Encoded Cascade-TniQ Complex.” Four primary structural models are included: the RNA-bound complex in both open and closed conformations of the Cas8 bundle, and the DNA-bound complex also exhibiting these two states. To facilitate visualization, a VMD state file is provided, allowing users to easily view the structural data.

For comparative purposes, we also include the previously published cryo-EM maps of the RNA-bound (EMD-20350) and DNA-bound (EMD-20351) Cascade-TniQ complexes, as reported by Halpin-Healy et al. in “Structural Basis of DNA Targeting by a Transposon-Encoded CRISPR–Cas System” (Nature 577, 271–274, 2020). These cryo-EM maps display low resolution in the Cas8 bundle region, underscoring the importance of the newly provided models. We advise the use of the software Chimera X for the visualization of the Cas8 bundle and other low-resolution regions in the Cryo-EM maps.

The provided files include:

• rna_hh_open.pdb: RNA-bound Cascade-TniQ complex in an open conformation of the Cas8 bundle.
• rna_hh_closed.pdb: RNA-bound Cascade-TniQ complex in a closed conformation of the Cas8 bundle.
• dna_hh_open.pdb: DNA-bound Cascade-TniQ complex in an open conformation of the Cas8 bundle.
• dna_hh_closed.pdb: DNA-bound Cascade-TniQ complex in a closed conformation of the Cas8 bundle.
• emd_20350.map: Cryo-EM map of the RNA-bound Cascade-TniQ complex (Halpin-Healy et al., Nature, 2020).
• emd_20351.map: Cryo-EM map of the DNA-bound Cascade-TniQ complex (Halpin-Healy et al., Nature, 2020).
• new-state-final-2.vmd: VMD state file for loading and visualizing the structural data, accessible through the command "vmd -e new-state-final-2.vmd". This state file applies the color schemes used in Patel et al.'s figures for consistency.