Nucleoid remodeling during environmental adaptation is regulated by HU-dependent DNA bundling.

Remesh SG, Verma SC, Chen JH, Ekman AA, Larabell CA, Adhya S, Hammel M, Nat Commun 11(1):2905 (2020) Europe PMC

SASDFS6 – DNA-binding protein HU-alpha, E34K mutant bound to 80 bp DNA (ratio DNA:Protein 1:2.5)

80bp_DNA Forward
80bp_DNA Reverse
DNA-binding protein HU-alpha, E34K
MWexperimental 290 kDa
MWexpected 316 kDa
VPorod 386 nm3
log I(s) 3.75×101 3.75×100 3.75×10-1 3.75×10-2
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E34K small angle scattering data  s, nm-1
ln I(s)
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E34K Guinier plot ln 3.76×101 Rg: 7.0 nm 0 (7.0 nm)-2 s2
(sRg)2I(s)/I(0)
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E34K Kratky plot 1.104 0 3 sRg
p(r)
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E34K pair distance distribution function Rg: 7.3 nm 0 Dmax: 25 nm

Data validation


Fits and models


log I(s)
 s, nm-1
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E34K CHIMERA model
80bp_DNA Forward 80bp_DNA Reverse DNA-binding protein HU-alpha, E34K CHIMERA model

Synchrotron SAXS data from solutions of DNA-binding protein HU-alpha, E34K mutant bound to 80 bp DNA (ratio DNA:Protein 1:2.5) in 20mM HEPES, 100mM NaCl, 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 Pilatus3 X 2M detector at a sample-detector distance of 1.5 m and at a wavelength of λ = 0.103 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). at 10°C. 300 successive 3 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.

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, E34K
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Other
Mon. MW   9.5 kDa
 
UniProt   P0ACF0 (1-90)
Sequence   FASTA