A new protein-dependent riboswitch activates ribosomal frameshifting

Betts J, Jeffries C, Passchier T, Kung H, Graham S, Abdelhamid M, Howard J, Craggs T, Graham S, Brierley I, Leake M, Quinn S, Hill C, (2025) DOI

SASDWS5 – 2A protein from Theiler's murine encephalomyelitis virus (TMEV) bound to stimulatory element RNA

Genome polyprotein
Stimulatory element RNA
MWexperimental 28 kDa
MWexpected 29 kDa
VPorod 46 nm3
log I(s) 6.64×103 6.64×102 6.64×101 6.64×100
Genome polyprotein Stimulatory element RNA small angle scattering data  s, nm-1
ln I(s)
Genome polyprotein Stimulatory element RNA Guinier plot ln 6.64×103 Rg: 2.2 nm 0 (2.2 nm)-2 s2
(sRg)2I(s)/I(0)
Genome polyprotein Stimulatory element RNA Kratky plot 1.104 0 3 sRg
p(r)
Genome polyprotein Stimulatory element RNA pair distance distribution function Rg: 2.2 nm 0 Dmax: 7.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Genome polyprotein Stimulatory element RNA CORAL model

log I(s)
 s, nm-1
Genome polyprotein Stimulatory element RNA MONSA model

Synchrotron SAXS data from solutions of 2A protein bound to stimulatory element RNA in 20 mM HEPES pH 7.4, 400 mM KCl 2% v/v glycerol, 1 mM DTT, were collected on the EMBL P12 beam line at PETRA III (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.123983 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 50.00 μl sample at 8.0 mg/ml was injected at a 0.35 ml/min flow rate onto a GE Superdex 75 Increase 5/150 column at 20°C. 41 successive 0.250 second frames were collected through the SEC-elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

The full entry archive contains the unsubtracted SEC-SAXS 1D-data frames in addition to MONSA and CORAL modelling routines.

Genome polyprotein (2A protein)
Mol. type   Protein
Organism   Theiler's murine encephalomyelitis virus (strain GDVII)
Olig. state   Monomer
Mon. MW   17.4 kDa
 
UniProt   Q88595 (923-1055)
Sequence   FASTA
 
Stimulatory element RNA
Mol. type   RNA
Organism   synthetic RNA
Olig. state   Monomer
Mon. MW   11.8 kDa
Sequence   FASTA