5 figures, 3 videos and 1 table

Figures

Figure 1 with 2 supplements
CryoEM structure of RecBCD/DNA complex at 3.8 Å resolution.

(A) Cartoon representation of the RecBCD complex within a transparent surface showing the electron density. The RecB subunit is in orange, RecC in cyan, RecD is in green, and the DNA is in pink, (B) …

https://doi.org/10.7554/eLife.18227.003
Figure 1—figure supplement 1
EM Data statistics.

(A) An example of an aligned and summed image stack showing particle distribution. (B) Selection of 2D class averages representing different particle views. (C) Final map coloured by local …

https://doi.org/10.7554/eLife.18227.004
Figure 1—figure supplement 2
Representative sections of electron density (contorted at 4 σ).

(A) Electron density corresponding to the bound DNA fork substrate, (B) Representative regions at the centre of the RecBCD complex to illustrate the quality of the maps, (C) The 2B/SH3 domain of the …

https://doi.org/10.7554/eLife.18227.005
Figure 2 with 2 supplements
Interactions with the 5’-tail and the RecD subunit.

Inset: location of the close ups shown in this figure relative to Figure 1. (A) Binding of the 5’-tail of the DNA fork across the RecD subunit, (B) Close up of the electron density (contoured at 4 σ)…

https://doi.org/10.7554/eLife.18227.006
Figure 2—figure supplement 1
Structural features of the RecB subunit.

Insets show location of the zoomed in regions. (A) Conformational changes of the 1A and 2A domains of RecB associated with nucleotide binding (initiation complex (Singleton et al., 2004) shown in …

https://doi.org/10.7554/eLife.18227.007
Figure 2—figure supplement 2
Nucleotide binding at the RecD subunit.

Surprisingly, only very weak electron density for bound nucleotide is visible at the ATP-binding site of RecD even though ADPNP is bound at the ATP-binding site of the RecB subunit. However, the …

https://doi.org/10.7554/eLife.18227.008
SH3 fold in RecD.

Inset: location of the close ups shown in this figure relative to Figure 1. (A) Contact regions of the SH3 domain (blue) with DNA (pink) and peptide (RecB, orange), (B) Cartoon representation of the …

https://doi.org/10.7554/eLife.18227.009
Figure 4 with 1 supplement
Conformational changes that result in nuclease activation.

Inset: location of the close ups shown in this figure. The series of conformational changes from the initiation complex (grey) begins in the top left corner. Supplementary Videos 13 show these …

https://doi.org/10.7554/eLife.18227.010
Figure 4—figure supplement 1
Electron density showing relocation of the linker peptide and nuclease blocking helix.

Inset shows location of the zoomed in region. (A) Electron density (contoured at 2.5 σ) showing lack of density for the nuclease blocking helix in the position seen in the initiation complex (grey). …

https://doi.org/10.7554/eLife.18227.011
Cartoon summarising the conformational changes that proceed from the initiation complex to nuclease activation.
https://doi.org/10.7554/eLife.18227.015

Videos

Video 1
Overall changes in the RecD subunit.
https://doi.org/10.7554/eLife.18227.012
Video 2
Overall changes in the RecC subunit.
https://doi.org/10.7554/eLife.18227.013
Video 3
Changes in the linker peptide region.
https://doi.org/10.7554/eLife.18227.014

Tables

Table 1

Refinement of the final model.

https://doi.org/10.7554/eLife.18227.002
Final particles74656
Resolution (Å)3.8
CC (whole map)0.838
CC (around atoms)0.875
R.m.s. deviations
Bond lengths (Å)0.01
Bond angles (°)1.04
Ramachandran
Favoured (%)94.1
Outliers8/2874
Rotamers
Favoured (%)99.1
Outliers1/2426

Download links