Structure of the PCBP2/stem-loop IV complex underlying translation initiation mediated by the poliovirus type I IRES.

Beckham SA, Matak MY, Belousoff MJ, Venugopal H, Shah N, Vankadari N, Elmlund H, Nguyen JHC, Semler BL, Wilce MCJ, Wilce JA, Nucleic Acids Res (2020) Europe PMC

SASDHA5 – Truncated poly(rC)-binding protein 2 (PCBP2-ΔKH1-KH2) with bound modified RNA stem loop IV poliovirus IRES (SLIVm)

modified stem loop IV poliovirus IRES, nucleotides 278-398
Truncated poly(rC)-binding protein 2 (ΔKH1-KH2)
MWexperimental 64 kDa
MWexpected 59 kDa
VPorod 126 nm3
log I(s) 2.05×10-2 2.05×10-3 2.05×10-4 2.05×10-5
modified stem loop IV poliovirus IRES, nucleotides 278-398 Truncated poly(rC)-binding protein 2 (ΔKH1-KH2) small angle scattering data  s, nm-1
ln I(s)
modified stem loop IV poliovirus IRES, nucleotides 278-398 Truncated poly(rC)-binding protein 2 (ΔKH1-KH2) Guinier plot ln 2.06×10-2 Rg: 3.5 nm 0 (3.5 nm)-2 s2
(sRg)2I(s)/I(0)
modified stem loop IV poliovirus IRES, nucleotides 278-398 Truncated poly(rC)-binding protein 2 (ΔKH1-KH2) Kratky plot 1.104 0 3 sRg
p(r)
modified stem loop IV poliovirus IRES, nucleotides 278-398 Truncated poly(rC)-binding protein 2 (ΔKH1-KH2) pair distance distribution function Rg: 3.6 nm 0 Dmax: 12.2 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of truncated poly(rC)-binding protein 2 (PCBP2-KH1-KH2) with bound modified RNA stem loop IV poliovirus IRES (SLIVm) in 5 mM HEPES-KOH, 25 mM KCl, 2 mM MgCl2, 2 mM DTT, 4 % glycerol, 0.1 mM EDTA, pH 7.5 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 2.7 m and at a wavelength of λ = 0.10322 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 100 μl sample was injected at a 0.40 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 15°C. 720 successive 5 second frames were collected through the SEC elution. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Sample injection concentration = UNKNOWN

modified stem loop IV poliovirus IRES, nucleotides 278-398 (SLIVm)
Mol. type   RNA
Organism   Human poliovirus 1 Mahoney
Olig. state   Monomer
Mon. MW   40.9 kDa
Sequence   FASTA
 
Truncated poly(rC)-binding protein 2 (ΔKH1-KH2) (PCBP2-(ΔKH1-KH2))
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   18.1 kDa
 
UniProt   Q15366 (11-168)
Sequence   FASTA