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

SASDGQ5 – Vibrio cholerae hexameric DnaB helicase with bound ATP (VcDnaB.ATP)

Replicative DNA helicase (DnaB)
MWexperimental 328 kDa
MWexpected 317 kDa
VPorod 585 nm3
log I(s) 2.78×10-1 2.78×10-2 2.78×10-3 2.78×10-4
Replicative DNA helicase (DnaB) small angle scattering data  s, nm-1
ln I(s)
Replicative DNA helicase (DnaB) Guinier plot ln 2.78×10-1 Rg: 4.6 nm 0 (4.6 nm)-2 s2
Replicative DNA helicase (DnaB) Kratky plot 1.104 0 3 sRg
Replicative DNA helicase (DnaB) pair distance distribution function Rg: 4.7 nm 0 Dmax: 13 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of the DnaB helicase hexamer in 20 mM Tris-HCl, 100 mM NaCl, 1 mM ATP, pH 8.8 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a Eiger 4M 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 48.00 μl sample at 28 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 15°C. 700 successive 2.990 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 10 identical frames obtained around the maximum of the elution peak were averaged and further processed.

Replicative DNA helicase (DnaB) (DnaB.ATP)
Mol. type   Protein
Organism   Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Olig. state   Hexamer
Mon. MW   52.8 kDa
UniProt   A0A0H3AIB6 (1-468)
Sequence   FASTA