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) PubMed

SASDAS8 – kLANA 1008-1150 -- kLBS1-2 complex 8:2 (partially dissociated)

kLBS1-2 DNA
ORF73 tetramer
ORF73 octamer
kLBS1-2 DNA two monomers
MWI(0) 150 kDa
MWexpected 65 kDa
VPorod 250 nm3
log I(s) 1.61×104 1.61×103 1.61×102 1.61×101
kLBS1-2 DNA ORF73 tetramer ORF73 octamer kLBS1-2 DNA two monomers small angle scattering data  s, nm-1
ln I(s)
kLBS1-2 DNA ORF73 tetramer ORF73 octamer kLBS1-2 DNA two monomers Guinier plot ln 1.62×104 Rg: 4.8 nm 0 (4.8 nm)-2 s2
(sRg)2I(s)/I(0)
kLBS1-2 DNA ORF73 tetramer ORF73 octamer kLBS1-2 DNA two monomers Kratky plot 1.104 0 3 sRg
p(r)
kLBS1-2 DNA ORF73 tetramer ORF73 octamer kLBS1-2 DNA two monomers pair distance distribution function Rg: 4.8 nm 0 Dmax: 16 nm

Data validation


Fits and models


log I(s)
 s, nm-1
kLBS1-2 DNA ORF73 tetramer ORF73 octamer kLBS1-2 DNA two monomers NONE model
kLBS1-2 DNA ORF73 tetramer ORF73 octamer kLBS1-2 DNA two monomers NONE model
kLBS1-2 DNA ORF73 tetramer ORF73 octamer kLBS1-2 DNA two monomers NONE model

Synchrotron SAXS data from solutions of kLANA 1008-1150 -- kLBS1-2 complex 8:2 (partially dissociated) in 25 mM Na/K Phosphate, pH 7.5 were collected on the EMBL P12 camera at the PETRA III storage ring (Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.12 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 0.40 mg/ml was measured at 10°C. 20 successive 0.050 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.

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

kLBS1-2 DNA
Mol. type   DNA
Organism   unidentified herpesvirus
Olig. state   Monomer
Mon. MW   24 kDa
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 octamer (kLANA)
Mol. type   Protein
Organism   Human herpesvirus 8
Olig. state   Octamer
Mon. MW   15.8 kDa
 
UniProt   Q9QR71 (1008-1150)
Sequence   FASTA
 
kLBS1-2 DNA two monomers
Mol. type   RNA
Organism   unidentified herpesvirus
Olig. state   Monomers
Mon. MW   24 kDa