Flexibility of the linker between the domains of DNA methyltransferase SsoII revealed by small-angle X-ray scattering: implications for transcription regulation in SsoII restriction-modification system.

Konarev PV, Kachalova GS, Ryazanova AY, Kubareva EA, Karyagina AS, Bartunik HD, Svergun DI, PLoS One 9(4):e93453 (2014) Europe PMC

SASDLT5 – Modification methylase SsoII (M.SsoII) protein bound to 12-bp DNA

Modification methylase SsoII
12-bp DNA
MWI(0) 44 kDa
MWexpected 51 kDa
VPorod 85 nm3
log I(s) 2.39×102 2.39×101 2.39×100 2.39×10-1
Modification methylase SsoII 12-bp DNA small angle scattering data  s, nm-1
ln I(s)
Modification methylase SsoII 12-bp DNA Guinier plot ln 2.39×102 Rg: 2.8 nm 0 (2.8 nm)-2 s2
(sRg)2I(s)/I(0)
Modification methylase SsoII 12-bp DNA Kratky plot 1.104 0 √3 sRg
p(r)
Modification methylase SsoII 12-bp DNA pair distance distribution function Rg: 2.8 nm 0 Dmax: 11 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Modification methylase SsoII (M.SsoII) protein bound to 12-bp DNA Rg histogram Rg, nm
Modification methylase SsoII 12-bp DNA EOM/RANCH model
Modification methylase SsoII 12-bp DNA EOM/RANCH model
Modification methylase SsoII 12-bp DNA EOM/RANCH model
Modification methylase SsoII 12-bp DNA EOM/RANCH model
Modification methylase SsoII 12-bp DNA EOM/RANCH model
log I(s)
 s, nm-1
Modification methylase SsoII 12-bp DNA MONSA model
Synchrotron SAXS data from solutions of M.SsoII protein bound to 12-bp DNA in 50 mM Na-phosphate buffer, pH 7 were collected on the EMBL X33 beam line at DORIS III (DESY, Hamburg, Germany) using a MAR 345 Image Plate detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 1.00 mg/ml was measured at 20°C. One 120 second frame was collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Tags: X33
Modification methylase SsoII (m.SsoII)
Mol. type   Protein
Organism   Shigella sonnei
Olig. state   Monomer
Mon. MW   42.9 kDa
 
UniProt   P34879 (1-379)
Sequence   FASTA
 
12-bp DNA (12-bp DNA)
Mol. type   DNA
Olig. state   Monomer
Mon. MW   7.7 kDa
Sequence   FASTA