Diving into the structural architecture of the B2 SINE ribozyme

Trushar Patel.

SASDV93 – B2 SINE ribozyme with B2J Mutation

B2 short interspaced nuclear element (SINE) RNA with two point mutations
MWexperimental 72 kDa
MWexpected 57 kDa
VPorod 136 nm3
log I(s) 2.71×10-2 2.71×10-3 2.71×10-4 2.71×10-5
B2 short interspaced nuclear element (SINE) RNA with two point mutations small angle scattering data  s, nm-1
ln I(s)
B2 short interspaced nuclear element (SINE) RNA with two point mutations Guinier plot ln 2.71×10-2 Rg: 5.5 nm 0 (5.5 nm)-2 s2
(sRg)2I(s)/I(0)
B2 short interspaced nuclear element (SINE) RNA with two point mutations Kratky plot 1.104 0 3 sRg
p(r)
B2 short interspaced nuclear element (SINE) RNA with two point mutations pair distance distribution function Rg: 5.6 nm 0 Dmax: 17.3 nm

Data validation


Fits and models


log I(s)
 s, nm-1
B2 short interspaced nuclear element (SINE) RNA with two point mutations DAMMIN model

log I(s)
 s, nm-1
B2 short interspaced nuclear element (SINE) RNA with two point mutations DAMMIN model

log I(s)
 s, nm-1
B2 short interspaced nuclear element (SINE) RNA with two point mutations DAMMIN model

Synchrotron SAXS data from solutions of B2 SINE ribozyme with deletion mutation 81-124 in 5 mM Tris-HCl, 10 mM NaCl, 0.01% NP-40, 0.02 mM EDTA, 0.2 mM DTT, 0.5 mM MgCl2 and 1.5 % Glycerol, pH 7.9 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 = 0.094 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. A sample at 1.6 mg/ml was injected into a Shodex 403KW-4F HPLC column at 15°C using a flow rate of 0.16 ml/min. 600 successive 3 second frames were collected. Eight successive 3 second frames were selected through the SEC elution peak of the sample. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the high-concentration high-angle data to yield the final composite scattering curve.

B2 short interspaced nuclear element (SINE) RNA with two point mutations (B2J)
Mol. type   RNA
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   57.2 kDa
Sequence   FASTA