README for Figure 5b.csv 
*** This file contains the raw data obtained on DNA, DNA-MvaT and DNA-MvaT-gp4 complexes using bridging assay experiments represented in Figure 5b of 

Article: Novel anti-repression mechanism of H-NS proteins by a phage protein 
Authors: Ben Bdira, Erkelens, Qin, Volkov, Lippas, Bowring, Boyle, Ubbink, Dove, Dame 
Journal: Nucleic Acids Research 

Corresponding authors: fredjbdira@gmail.com and rtdame@chem.leidenuniv.nl 

Legend Figure 5: Validation of gp4 anti-bridging mechanism by point mutagenesis. (a) electrostatic surface of
gp4 WT and gp4 K42E/K45 mutant. (b) Inhibitory effects of gp4 mutant (red bars) and gp4 (black bars) on
MvaT DNA bridging activity at 0.9 and 1.2 ?M of gp4 using a 2.4 ?M concentration of MvaT (grey bars).
(c) ITC of MvaT2 with gp K42E/K45E. The red line shows the best fit of the data. (d) Analysis of the NMR
titration data of 15N MvaT2 with gp4 K42E/K45E. The upper panel depicts the 15N-1H TROSY spectra peak
intensities ratio of MvaT2 in the presence of 1.2 molar ratio of gp4 (I) and the free state (I0) versus the
protein residue number. Note the two-fold increase in the peak intensities of the DBD-linker region (shaded
in grey) due to the loss of the intramolecular electrostatic interactions between the DBD-linker -NTD within
MvaT2 subunits (18) and the intermolecular interactions with gp4. Lower panel shows weighted average
CSP of the MvaT resonances between the same points of titration. Resonances with CSP more than two
(orange line) or one (yellow line) standard deviation (SD) from the 10% trimmed mean (green dashed line)
are labelled and shown in orange, yellow and green bars, respectively. The blue and green bars are for
resonances with a reduction in peaks intensities and CSP higher or less than the 10% trimmed mean,
respectively.

*** The data were obtained using bridging assay experiments as described in the associated article. 

***The data labeled overview represents the values plotted in the graph of figure 5b. The raw data contains the individual replicates.
Column A: Sample number
Column B: Presence of bait DNA in sample
Column C: Presence of 32P-labeled prey DNA in sample
Column D: Amount of MvaT added to sample in micromolar
Column E: Amount of gp4 added to sample in micromolar
Column F: Counts per minutes 
Column G: DNA recovery (%)

Columns B, C, D and E indicate the contents of the sample. Column F indicates the radioactivity of the sample. Column G indicates the DNA recovery, which is calculated by subtracting the value in column F of the control (sample minus bait DNA) from the sample itself. This value is then divided by the value in column F of sample 1 and multiplied by 100%. 









