Structural basis for topological regulation of Tn3 resolvase.

Montaño SP, Rowland SJ, Fuller JR, Burke ME, MacDonald AI, Boocock MR, Stark WM, Rice PA, Nucleic Acids Res (2022) Europe PMC

SASDNH7 – Tn3 resolvase dimer bound to site II DNA sequence

Transposon Tn3 resolvase
Tn3 res, resolvase binding site II
MWexperimental 73 kDa
MWexpected 75 kDa
VPorod 217 nm3
log I(s) 5.60×101 5.60×100 5.60×10-1 5.60×10-2
Transposon Tn3 resolvase Tn3 res, resolvase binding site II small angle scattering data  s, nm-1
ln I(s)
Transposon Tn3 resolvase Tn3 res, resolvase binding site II Guinier plot ln 5.60×101 Rg: 5.0 nm 0 (5.0 nm)-2 s2
(sRg)2I(s)/I(0)
Transposon Tn3 resolvase Tn3 res, resolvase binding site II Kratky plot 1.104 0 √3 sRg
p(r)
Transposon Tn3 resolvase Tn3 res, resolvase binding site II pair distance distribution function Rg: 4.9 nm 0 Dmax: 14.9 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Transposon Tn3 resolvase Tn3 res, resolvase binding site II CUSTOM IN-HOUSE model
Synchrotron SAXS data from solutions of Tn3 resolvase dimer bound to a site II DNA sequence in 25 mM Tris, 150 mM NaCl, 10 mM MgCl2, 1% glycerol, pH 7.5 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS), Argonne National Laboratory (Lemont, IL, USA) using a Pilatus 100K detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.103 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 500.00 μl sample at 3 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 10/300 column at 22°C. 800 successive 1.090 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.

Transposon Tn3 resolvase (Tn3 resolvase)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Dimer
Mon. MW   21.5 kDa
 
UniProt   P0ADI2 (1-185)
Sequence   FASTA
 
Tn3 res, resolvase binding site II (Site II)
Mol. type   DNA
Olig. state   Monomer
Mon. MW   32.1 kDa
Sequence   FASTA