Structural analysis of monomeric retroviral reverse transcriptase in complex with an RNA/DNA hybrid.

Nowak E, Potrzebowski W, Konarev PV, Rausch JW, Bona MK, Svergun DI, Bujnicki JM, Le Grice SF, Nowotny M, Nucleic Acids Res 41(6):3874-87 (2013) PubMed

SASDAW5 – XMRV RT + DNA/RNA hybrid

apo XMRV RT
RNA_DNA hybrid substrate
MWexperimental 91 kDa
MWexpected 90 kDa
VPorod 155 nm3
log I(s) 7.83×101 7.83×100 7.83×10-1 7.83×10-2
apo XMRV RT RNA_DNA hybrid substrate small angle scattering data  s, nm-1
ln I(s)
apo XMRV RT RNA_DNA hybrid substrate Guinier plot ln 7.83×101 Rg: 3.5 nm 0 (3.5 nm)-2 s2
(sRg)2I(s)/I(0)
apo XMRV RT RNA_DNA hybrid substrate Kratky plot 1.104 0 3 sRg
p(r)
apo XMRV RT RNA_DNA hybrid substrate pair distance distribution function Rg: 3.6 nm 0 Dmax: 11.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
apo XMRV RT RNA_DNA hybrid substrate CRYSOL model

Synchrotron SAXS data from solutions of XMRV RT + DNA/RNA hybrid in 10 mM HEPES 100 mM KCl 5% Glycerol, pH 6.5 were collected on the EMBL X33 camera at the DORIS III storage ring (Hamburg, Germany) using a Pilatus 1M-W detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.15 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). at 10°C. Eight successive 15 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.

apo XMRV RT (XM)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   75 kDa
Sequence   FASTA
 
RNA_DNA hybrid substrate (PPT18)
Mol. type   Other
Olig. state   Monomer
Mon. MW   15 kDa
Sequence   FASTA