Nucleoid remodeling during environmental adaptation is regulated by HU-dependent DNA bundling (supplementary)

Soumya G Remesh.

SASDFZ6 – DNA-binding protein HU-alpha, E38K/V42L double mutant bound to 80 bp DNA (ratio DNA:Protein 1:16)

80bp_DNA Forward
80bp_DNA Reverse
DNA-binding protein HU-alpha, E38K/V42L double mutant
MWexperimental 330 kDa
MWexpected 202 kDa
VPorod 401 nm3
log I(s) 8.88×102 8.88×101 8.88×100 8.88×10-1
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E38K/V42L double mutant small angle scattering data  s, nm-1
ln I(s)
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E38K/V42L double mutant Guinier plot ln 8.88×102 Rg: 6.3 nm 0 (6.3 nm)-2 s2
(sRg)2I(s)/I(0)
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E38K/V42L double mutant Kratky plot 1.104 0 3 sRg
p(r)
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E38K/V42L double mutant pair distance distribution function Rg: 7.5 nm 0 Dmax: 27.3 nm

Data validation


Fits and models


log I(s)
 s, nm-1
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E38K/V42L double mutant CHIMERA model

Synchrotron SAXS data from solutions of DNA-binding protein HU-alpha, E38K/V42L double mutant bound to 80 bp DNA (ratio DNA:Protein 1:16) in 50 mM Tris-HCl, 150 mM NaCl, 1 mM DTT, 1 mM PMSF, pH 7.5 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS) storage ring (Berkeley, CA, USA) using a MAR 165 CCD detector at a sample-detector distance of 1.5 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). at 10°C. 32 successive 0.100 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Storage temperature = UNKNOWN. Concentration = UNKNOWN

80bp_DNA Forward
Mol. type   DNA
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   24.8 kDa
Sequence   FASTA
 
80bp_DNA Reverse
Mol. type   DNA
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   24.6 kDa
Sequence   FASTA
 
DNA-binding protein HU-alpha, E38K/V42L double mutant
Mol. type   Protein
Organism   Linked to wild-type UniProt entry P0ACF0 from E coli
Olig. state   16-mer
Mon. MW   9.5 kDa
Sequence   FASTA