Studies of bacteriophage Mu transposition paved the way for understanding retroviral integration and V(D)J recombination as well as many other DNA transposition reactions. Here we report the structure of the Mu transpososome—Mu transposase (MuA) in complex with bacteriophage DNA ends and target DNA—determined from data that extend anisotropically to 5.2 Å, 5.2 Å and 3.7 Å resolution, in conjunction with previously determined structures of individual domains. The highly intertwined structure illustrates why chemical activity depends on formation of the synaptic complex, and reveals that individual domains have different roles when bound to different sites. The structure also provides explanations for the increased stability of the final product complex and for its preferential recognition by the ATP-dependent unfoldase ClpX. Although MuA and many other recombinases share a structurally conserved ‘DDE’ catalytic domain, comparisons among the limited set of available complex structures indicate that some conserved features, such as catalysis in trans and target DNA bending, arose through convergent evolution because they are important for function.
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- Video 1: Ribbon drawing of the transpososome structure rotating 360° (4,785 KB, Download)
- The complex is rotating about the crystallographic twofold axis that relates the red and blue halves. Colours are as in the main text: bacteriophage Mu end DNAs are red and blue, target DNA black, and the scissile phosphate and active site residues are yellow. The darker-colored subunits catalyze DNA cleavage and strand transfer and the lighter-colored subunits aid in complex assembly and stability.
- Video 2: Closeup view of the experimental electron density, after improvement with Parrot, and contoured at 1.3 and 2.3 Sigma, rotating 360° (9,536 KB, Download)
- The rotation axis and colors are as in the main text and Supplementary Video 1.
- Supplementary Information (1.1 MB)
This file contains Supplementary Figures 1-4, Supplementary References and Supplementary Table 1.