Contrasting DNA-binding behaviour by ISL1 and LHX3 underpins differential gene targeting in neuronal cell specification

Smith N, Wilkinson-White L, Kwan A, Trewhella J, Matthews J, Journal of Structural Biology: X :100043 (2020) DOI

SASDJC9 – Insulin gene enhancer protein Isl-1 LIM-interaction domain (Isl1-LID) fused to the LIM/homeobox protein Lhx3 LIM-homeodomain (Lhx3-LIMHD): LLHD3

LIM/homeobox protein Lhx3
Insulin gene enhancer protein ISL-1
MWI(0) 29 kDa
MWexpected 27 kDa
VPorod 39 nm3
log I(s) 7.35×10-3 7.35×10-4 7.35×10-5 7.35×10-6
LIM/homeobox protein Lhx3 Insulin gene enhancer protein ISL-1 small angle scattering data  s, nm-1
ln I(s)
LIM/homeobox protein Lhx3 Insulin gene enhancer protein ISL-1 Guinier plot ln 7.36×10-3 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
LIM/homeobox protein Lhx3 Insulin gene enhancer protein ISL-1 Kratky plot 1.104 0 √3 sRg
p(r)
LIM/homeobox protein Lhx3 Insulin gene enhancer protein ISL-1 pair distance distribution function Rg: 3.5 nm 0 Dmax: 12 nm

Data validation

Fits and models

log I(s)
 s, nm-1
LIM/homeobox protein Lhx3 Insulin gene enhancer protein ISL-1 MONSA model
log I(s)
 s, nm-1
LIM/homeobox protein Lhx3 Insulin gene enhancer protein ISL-1 DAMMIN model
Synchrotron SAXS data from solutions of LLHD3 (Isl1-LID fused to Lhx3-LIMHD) in 20 mM sodium phosphate buffer, 100 mM NaCl, 1 mM DTT, pH 7.4 were collected on the SAXS/WAXS beam line at the Australian Synchrotron (Melbourne, Australia) using a Pilatus 1M detector at a sample-detector distance of 0.9 m and at a wavelength of λ = 0.108 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 0.33 mg/ml was measured at 27°C. 32 successive 1 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.

Storage temperature = UNKNOWN

LIM/homeobox protein Lhx3 (Lhx3)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   23.2 kDa
 
UniProt   P50481 (28-227)
Sequence   FASTA
 
Insulin gene enhancer protein ISL-1 (Isl1)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   4.2 kDa
 
UniProt   P61372 (262-291)
Sequence   FASTA