The herpes viral transcription factor ICP4 forms a novel DNA recognition complex

Tunnicliffe R, Lockhart-Cairns M, Levy C, Mould A, Jowitt T, Sito H, Baldock C, Sandri-Goldin R, Golovanov A, Nucleic Acids Research 2017 May 13 DOI

SASDB48 – Major viral transcription factor ICP4/Immediate Early 3 DNA (ICP4N IE3_19mer) complex

Major viral transcription factor ICP4
Immediate Early 3
MWexperimental 60.4 kDa
MWexpected 60 kDa
VPorod 89.7 nm3
log I(s) 1.14×102 1.14×101 1.14×100 1.14×10-1
Major viral transcription factor ICP4 Immediate Early 3 small angle scattering data  s, nm-1
ln I(s)
Major viral transcription factor ICP4 Immediate Early 3 Guinier plot ln 1.14×102 Rg: 2.5 nm 0 (2.5 nm)-2 s2
(sRg)2I(s)/I(0)
Major viral transcription factor ICP4 Immediate Early 3 Kratky plot 1.104 0 3 sRg
p(r)
Major viral transcription factor ICP4 Immediate Early 3 pair distance distribution function Rg: 2.5 nm 0 Dmax: 8.3 nm

Experimental data validation


Fits and models


log I(s)
 s, nm-1
Major viral transcription factor ICP4 Immediate Early 3 NONE model
log I(s)
 s, nm-1
Major viral transcription factor ICP4 Immediate Early 3 DAMFILT model
Synchrotron SAXS data from solutions of Major viral transcription factor ICP4/Immediate Early 3 DNA (ICP4N IE3_19mer) complex in 20 mM HEPES, 150 mM NaCl, pH 7.4 were collected using HPLC SEC-SAXS at the B21 beam line at the Diamond Light Source (Oxfordshire, UK) using a Pilatus 2M detector at a sample-detector distance of 3.9 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4π sin θ/λ and 2θ is the scattering angle). The sample was separated using a Shodex KW-403 SEC column and Agilent HPLC before exposure to X-rays to isolate the ICP4N IE3_19mer complex from any dissociated monomer. 50 µL of ICP4N•IE3_19mer complex was loaded onto the Shodex column and the eluent was flowed through the SAXS beam at 0.15 mL/min; the buffer used as the background was collected after one SEC column volume. Data were collected at 20°C at one second intervals.

Major viral transcription factor ICP4 (ICP4N)
Mol. type   Protein
Organism   Human herpesvirus 1 (strain 17)
Olig. state   Dimer
Mon. MW   24.3 kDa
 
UniProt   P08392 (258-487)
Sequence   FASTA
 
Immediate Early 3 (IE3)
Mol. type   DNA
Olig. state   Dimer
Mon. MW   5.8 kDa
Sequence   FASTA
 
PDB code   5MHK