Fig. 4: The cryo-EM structure of the LE-LE Hermes transpososome.

a Example of selected 2D classes of the transpososome from the data processing in RELION^74,75,76. The transposase dimers (H^d) are marked with white arrowheads and the DNAs are indicated with blue arrowheads. The box size is 280 Å. b Cryo-EM map of the transpososome composed of three Hermes dimers (A, B, and C) and of two LE-DNAs (6.3 Å resolution). c Cryo-EM map of the transpososome composed of four Hermes dimers (A, B, C, and D) and of two LE-DNAs (5.5 Å resolution). d Multi-body refinement composite map of the transpososome. Body 1 corresponds to the DNAs, the Hermes dimers A and C and the BED domains of dimer B (4.6 Å resolution). Body 2 and Body 3 correspond to Hermes dimers B and D, and were refined in two other maps at 10.2 and 10.9 Å resolution, respectively. e Model of the LE-LE Hermes transpososome inside the composite cryo-EM map. f, g The atomic model of the core of the LE-LE Hermes transpososome (PDB: 8EDG, this publication). g Left: the antiparallel LE-DNAs inside the complex (only the silhouette of the atomic surface of the Hermes dimers is shown). The features of the DNAs (TIR and STRs) are highlighted. Right: the atomic surface of the Hermes dimers is displayed while the LE-DNA 1 is shown in cartoon mode to emphasize the extended protein/DNA interaction inside the transpososome. All the maps presented were sharpened and denoised with DeepEMhancer⁸⁵. The corresponding RELION’s post-processed maps are presented in Supplementary Figs. 5–8.