Conformational Changes of RORĪ³ During Response Element Recognition and Coregulator Engagement

Strutzenberg T, Zhu Y, Novick S, Garcia-Ordonez R, Doebelin C, He Y, Ra Chang M, Kamenecka T, Edwards D, Griffin P, Journal of Molecular Biology :167258 (2021) DOI

SASDLF5 – RORg2 bound to a Variant-RORgamma Response Element

Retinoid-related orphan receptor-gamma
Variant-RORgamma Response Element
MWI(0) 67 kDa
MWexpected 74 kDa
VPorod 112 nm3
log I(s) 1.26×101 1.26×100 1.26×10-1 1.26×10-2
Retinoid-related orphan receptor-gamma Variant-RORgamma Response Element small angle scattering data  s, nm-1
ln I(s)
Retinoid-related orphan receptor-gamma Variant-RORgamma Response Element Guinier plot ln 1.27×101 Rg: 4.4 nm 0 (4.4 nm)-2 s2
(sRg)2I(s)/I(0)
Retinoid-related orphan receptor-gamma Variant-RORgamma Response Element Kratky plot 1.104 0 √3 sRg
p(r)
Retinoid-related orphan receptor-gamma Variant-RORgamma Response Element pair distance distribution function Rg: 5.2 nm 0 Dmax: 22.1 nm

Data validation

Fits and models

log I(s)
 s, nm-1
RORg2 bound to a Variant-RORgamma Response Element Rg histogram Rg, nm
Synchrotron SAXS data from solutions of RORg2 bound to a Variant-RORgamma Response Element in 25 mM HEPES, 150 mM TCEP, 2% Glycerol, 5 mM DTT, 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 2 m and at a wavelength of λ = 0.127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.00 mg/ml was measured at 10°C. 33 successive 0.300 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.

Retinoid-related orphan receptor-gamma (RORg)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   55.9 kDa
 
UniProt   P51449 (25-518)
Sequence   FASTA
 
Variant-RORgamma Response Element (vRORE)
Mol. type   DNA
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   9.2 kDa
Sequence   FASTA