Study of the DnaB:DciA interplay reveals insights into the primary mode of loading of the bacterial replicative helicase.

Marsin S, Adam Y, Cargemel C, Andreani J, Baconnais S, Legrand P, Li de la Sierra-Gallay I, Humbert A, Aumont-Nicaise M, Velours C, Ochsenbein F, Durand D, Le Cam E, Walbott H, Possoz C, Quevillon-Cheruel S, Ferat JL, Nucleic Acids Res (2021) Europe PMC

SASDGP5 – Vibrio cholerae DciA

DciA
MWexperimental 18 kDa
MWexpected 18 kDa
VPorod 26 nm3
log I(s) 5.90×10-3 5.90×10-4 5.90×10-5 5.90×10-6
DciA small angle scattering data  s, nm-1
ln I(s)
DciA Guinier plot ln 5.90×10-3 Rg: 2.7 nm 0 (2.7 nm)-2 s2
(sRg)2I(s)/I(0)
DciA Kratky plot 1.104 0 √3 sRg
p(r)
DciA pair distance distribution function Rg: 2.8 nm 0 Dmax: 10.1 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of DciA in 20 mM Tris-HCl, 100 mM NaCl, pH 7.5 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a AVIEX PCCD170170 detector at a sample-detector distance of 1.8 m and at a wavelength of λ = 0.1033 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 95.00 μl sample at 5.7 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superose 6 10/300 column at 15°C. 254 successive 2 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.

The scattered intensities were displayed on an absolute scale (cm-1) using the scattering of water. Frames were examined individually using the US-SOMO HPLC module and 26 identical frames obtained around the maximum of the elution peak were averaged and further processed.

DciA (DciA)
Mol. type   Protein
Organism   Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Olig. state   Monomer
Mon. MW   18.4 kDa
 
UniProt   Q9KPH3 (1-157)
Sequence   FASTA