Integrative structural analysis of NF45-NF90 heterodimers reveals architectural rearrangements and oligomerization on binding dsRNA.

Winterbourne S, Jayachandran U, Zou J, Rappsilber J, Granneman S, Cook AG, Nucleic Acids Res 53(6) (2025) Europe PMC

SASDUA5 – Double-stranded RNA-binding domains of Interleukin enhancer-binding factor 3 (395-592)

Interleukin enhancer-binding factor 3
MWexperimental 23 kDa
MWexpected 21 kDa
VPorod 39 nm3
log I(s) 3.47×10-2 3.47×10-3 3.47×10-4 3.47×10-5
Interleukin enhancer-binding factor 3 small angle scattering data  s, nm-1
ln I(s)
Interleukin enhancer-binding factor 3 Guinier plot ln 3.47×10-2 Rg: 3.5 nm 0 (3.5 nm)-2 s2
(sRg)2I(s)/I(0)
Interleukin enhancer-binding factor 3 Kratky plot 1.104 0 √3 sRg
p(r)
Interleukin enhancer-binding factor 3 pair distance distribution function Rg: 3.5 nm 0 Dmax: 12.6 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Interleukin enhancer-binding factor 3 GASBOR model
Synchrotron SAXS data from solutions of Double-stranded RNA-binding domains of Interleukin enhancer-binding factor 3 (395-592) in 20 mM HEPES, 150 mM NaCl, 1 mM DTT, pH 7.5 were collected on the B21 beam line at the Diamond Light Source storage ring (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.0954 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 60.00 μl sample at 10 mg/ml was injected at a 0.10 ml/min flow rate onto a Cytiva Superdex 200 Increase 3.2/300 column at 15°C. 915 successive 0.005 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.

Interleukin enhancer-binding factor 3 (ILF3 dsRBDs)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   21.1 kDa
 
UniProt   Q9Z1X4 (395-592)
Sequence   FASTA