Increased association between Epstein-Barr virus EBNA2 from type 2 strains and the transcriptional repressor BS69 restricts B cell growth

Ponnusamy R, Khatri R, Correia P, Mancini E, Farrell P, West M, (2018) DOI

SASDEJ6 – Complex of Epstein-Barr nuclear antigen 2 (EBNA2 Type1, amino acids 381-455) and BS69 (Zinc finger MYND domain-containing protein 11: CC-MYND, amino acids 480-602)

Epstein-Barr nuclear antigen 2
Zinc finger MYND domain-containing protein 11
MWexperimental 69 kDa
MWexpected 76 kDa
VPorod 72 nm3
log I(s) 1.07×10-2 1.07×10-3 1.07×10-4 1.07×10-5
Epstein-Barr nuclear antigen 2 Zinc finger MYND domain-containing protein 11 small angle scattering data  s, nm-1
ln I(s)
Epstein-Barr nuclear antigen 2 Zinc finger MYND domain-containing protein 11 Guinier plot ln 1.07×10-2 Rg: 4.6 nm 0 (4.6 nm)-2 s2
(sRg)2I(s)/I(0)
Epstein-Barr nuclear antigen 2 Zinc finger MYND domain-containing protein 11 Kratky plot 1.104 0 √3 sRg
p(r)
Epstein-Barr nuclear antigen 2 Zinc finger MYND domain-containing protein 11 pair distance distribution function Rg: 4.7 nm 0 Dmax: 13.8 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Epstein-Barr nuclear antigen 2 Zinc finger MYND domain-containing protein 11 DAMMIN model
Epstein-Barr nuclear antigen 2 Zinc finger MYND domain-containing protein 11 DAMMIF model
Synchrotron SAXS data from solutions of Epstein-Barr nuclear antigen 2 (EBNA2) bound to BS69 CC-MYND in 20mM Tris-HCl, 100mM NaCl, 2% Sucrose and 1mM TCEP, pH 7.5 were collected on the B21 beam line at the Diamond Light Source storage ring (Oxfordshire, UK) using a Pilatus 2M detector at a sample-detector distance of 4.0 m and at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). The sample was measured using size exclusion chromatography SAXS (SEC-SAXS) at 25°C. 25 successive 3 second frames were collected through the SEC sample elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged and the scattering of an appropriate solvent-blank was subtracted.

SEC-SAXS was performed using the following parameters: Column type: S200 3.2/300 increase; Flow rate: 0.075 ml/min; Injection volume: 45 µl; Sample injection concentration: 9.0 mg/ml. The molar ratio of the injected sample components was 1:3 (EBNA2:BS69). The final complex contains four BS69 CC-MYND (2 dimers) with two EBNA2 Type1 (381-455) polypeptides.

Epstein-Barr nuclear antigen 2 (EBNA2 Type1)
Mol. type   Protein
Organism   Human gammaherpesvirus 4
Olig. state   Dimer
Mon. MW   7.8 kDa
 
UniProt   P12978 (381-455)
Sequence   FASTA
 
Zinc finger MYND domain-containing protein 11 (BS69 CC-MYND)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   15.1 kDa
 
UniProt   Q15326-1 (480-602)
Sequence   FASTA