KSHV but not MHV-68 LANA induces a strong bend upon binding to terminal repeat viral DNA.

Ponnusamy R, Petoukhov MV, Correia B, Custodio TF, Juillard F, Tan M, Pires de Miranda M, Carrondo MA, Simas JP, Kaye KM, Svergun DI, McVey CE, Nucleic Acids Res 43(20):10039-54 (2015) Europe PMC

SASDAQ8 – kLANA mutant dimer-tetramer mixture

ORF73 tetramer
ORF73 dimer
MWI(0) 30 kDa
MWexpected 47 kDa
VPorod 50 nm3
log I(s) 3.50×101 3.50×100 3.50×10-1 3.50×10-2
ORF73 tetramer ORF73 dimer small angle scattering data  s, nm-1
ln I(s)
ORF73 tetramer ORF73 dimer Guinier plot ln 3.50×101 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
ORF73 tetramer ORF73 dimer Kratky plot 1.104 0 3 sRg
p(r)
ORF73 tetramer ORF73 dimer pair distance distribution function Rg: 2.4 nm 0 Dmax: 9.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
ORF73 tetramer ORF73 dimer NONE model
ORF73 tetramer ORF73 dimer NONE model

Synchrotron SAXS data from solutions of kLANA mutant dimer-tetramer mixture in 25 mM Na/K Phosphate, pH 7.5 were collected on the BM29 beam line at the ESRF storage ring (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.4 m and at a wavelength of λ = 0.93 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.50 mg/ml was measured. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Study of oligomeric states of mLANA and kLANA and their complexes with DNA

ORF73 tetramer (kLANA mutant K1109E)
Mol. type   Protein
Organism   Human herpesvirus 8
Olig. state   Tetramer
Mon. MW   15.8 kDa
 
UniProt   Q9QR71 (1008-1150)
Sequence   FASTA
 
ORF73 dimer (kLANA mutant K1109E)
Mol. type   Protein
Organism   Human herpesvirus 8
Olig. state   Dimer
Mon. MW   15.8 kDa
 
UniProt   Q9QR71 (1008-1150)
Sequence   FASTA