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The red line represents the best fit of a Gaussian curve to the experimental data. The red dashed line shows the calculatedmaximum amplitude of the Gaussian curve. (**B**) Model with homo-dimers of CMPPQCC (WT) and CMPNPQCC(mutant). The red line represents the best fit of a Gaussian curve to the experimental data. The red dashed line shows thecalculated maximum amplitude of the Gaussian curve. (**C**) Model with hetero-dimers of CMPPQCC (WT) and WT (redline) and CMPNPQCC (mutant) and mutant (blue line). The black dashed line shows the calculated maximum amplitude of theGaussian curve.](gku320fig5){#F5} Most importantly, in contrast to the topology prediction, each subunit of the dimer would
be located on the opposite side of the helix (see 'Materials and Methods' section). Such a structure would only be allowed if thetwo subunits of the dimer were associated through an inter-subunit interface located on the bottom edge of the C-terminal helix(Figure [5B](#F5){ref-type="fig"}), which is consistent with the presence of the nucleotide analogues in the cleft between thetwo subunits. DISCUSSION {#SEC4} ========== Here, we report the first crystal structure of the DNA non-specifictopoisomerase I from the hyperthermophilic archaeon *Pyrococcus furiosus*, which is the first bacterial topo I structurereported to date. The structure presents some striking features. First, the mon f678ea9f9e
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