README for Figure1e.csv 
*** This file contains the raw data obtained on DNA, DNA-MvaT and DNA-gp4 complexes using tethered particle motion represented in Figure 1e 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 1:Effect of LUZ24 gp4 protein on P. aeruginosa growth and on the DNA binding modes of
MvaT. (a) Fold topology of MvaT monomer and protomer. The upper panel is a schematic representation
of MvaT monomer fold organization. In the lower panel is the structural model of the MvaT protomer
adopted from (18). The DBD are colored in yellow, site 1 in magenta, site 2 in cyan and the linker in blue.
(b) Heterologous expression of gp4 in P. aeruginosa wild-type cells and cells lacking either mvaU or mvaT.
The left panel shows serial dilutions (d) of cells growth in the absence of inducer and the right panel is in
the presence of Rhamnose for induction of gp4 expression. (c) Electrophoretic mobility-shift assay (EMSA) of 200 bp DNA substrate (32% GC) by 2 μM of MvaT wild type and MvaT2 in the presence of different
concentrations of gp4. (M) is for a 1kb DNA marker and (C) is for the 200 bp DNA substrate without
proteins. (d) NMR titration of 15N MvaT2:gp4 complex (1:1.2 molar ratio) with a 20 bp DNA substrate.
Representative chemical shift perturbations (CSP) of the MvaT2 DBD are shown and the corresponded
amino acid residues are depicted on the MvaT2 structure in (a). Black arrows indicate the directions of the
CSP. (e) The blue curve represents the RMS of DNA bound by MvaT at concentrations from 0 to 1200 nM
measured by the Tethered Particle Motion in the absence of gp4. The red curve is for the RMS of DNA
titrated by increased concentrations of gp4 between 0 and 2000 nM, in the absence of MvaT. The error bars
represent the standard deviation propagated from two independent measurements (each including ~100
individual DNA tethers); some error bars are hidden behind the data points. (f) & (g) Effects of gp4 on
MvaT DNA stiffening and bridging activities, respectively. Schematic representations of the TPM and
bridging assays are depicted in figure (S1b,c). The error bars are for the standard deviation from duplicate
experiments.

 *** The data were obtained using Tethered particle motion as described in the associated article. 

*** Data obtained for each protein concentration(s) is given in this table as plotted in figure 1e.
Column A: MvaT(nM) = Concentration of MvaT in nanomolar
Column B: gp4 (nM) = Concentration of gp4 in nanomolar
Column C: Replicate = number of replicate
Column D: RMS (nm) = Root mean square deviation in nanometer measured for each bead 
Column E: rho = anisotropic ratio (movement in x-direction/movement in y-direction) 
Column F: stdev = standard deviation 
Column G: FoV = field of view (data with the same FoV were collected in one image) 
Column H: bead = beads in each FoV are represented by a unique number 
The values given in column E and F are tether quality criteria and used for data filtering as described in the associated article.
