Structural analysis of the Candida albicans mitochondrial DNA maintenance factor Gcf1p reveals a dynamic DNA-bridging mechanism.

Tarrés-Solé A, Battistini F, Gerhold JM, Piétrement O, Martínez-García B, Ruiz-López E, Lyonnais S, Bernadó P, Roca J, Orozco M, Le Cam E, Sedman J, Solà M, Nucleic Acids Res (2023) Europe PMC

SASDP36 – N-terminal truncated Gcf1p (amino acids 59-245) bound to DNA

Af2_20 DNA
Gcf1p(Δ58)
MWexperimental 33 kDa
MWexpected 35 kDa
VPorod 52 nm3
log I(s) 1.56×10-1 1.56×10-2 1.56×10-3 1.56×10-4
Af2_20 DNA Gcf1p(Δ58) small angle scattering data  s, nm-1
ln I(s)
Af2_20 DNA Gcf1p(Δ58) Guinier plot ln 1.56×10-1 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Af2_20 DNA Gcf1p(Δ58) Kratky plot 1.104 0 √3 sRg
Dmax: 10.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Af2_20 DNA Gcf1p(Δ58) OTHER model
Synchrotron SAXS data from solutions of truncated Gcf1p (amino acids 59-245) bound to DNA in 25 mM Tris, 20 mM NaCl, pH 8 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Eiger 4M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.3 and 2 mg/ml were measured at 10°C. 40 successive 0.045 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. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Af2_20 DNA
Mol. type   DNA
Olig. state   Monomer
Mon. MW   12.4 kDa
Sequence   FASTA
 
Gcf1p(Δ58)
Mol. type   Protein
Organism   Candida albicans (strain SC5314 / ATCC MYA-2876)
Olig. state   Monomer
Mon. MW   22.5 kDa
 
UniProt   Q59QB8 (59-245)
Sequence   FASTA