Human RAD51 post-synaptic complexes mutant (V273P, D274G)

PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER

Identification of fidelity-governing factors in human recombinases DMC1 and RAD51 from cryo-EM structures.

25 mM Tris-HCl, pH 7.5, 50 mM KCl and 1 mM dithiothreitol) containing 2 mM AMP-PNP and 5 mM CaCl2

RAD51-dsDNA filament (V273P D274G mutant)

Filaments were manually picked, and segments were extracted using a box size of 384 pixel and an inter-box distance of ~10% of the box length.

protein sample were applied onto a pre-glow-discharged graphene-oxide coated Quantifoil holey carbon grid (1.2/1.3, 200 mesh) using published protocol

The grids were blotted for 1 sec at 22 degree C with 100% relative humidity and plunge-frozen in liquid ethane cooled by liquid nitrogen using a Vitrobot Mark IV (Thermo Fisher).

Authors know the sequence of chains D/E. Chain D: TCA GCT GTT GCC CGT. Chain E: ACG GGC AAC AGC TGA. Since the DNA sequence could not be revealed by the helical reconstruction method, authors have used poly-T for chain D, poly-A for chain E respectively for model building.
DNA (5'-D(P*TP*TP*TP*TP*TP*TP*TP*TP*T)-3')

DNA (5'-D(P*AP*AP*AP*AP*AP*AP*AP*AP*A)-3')

scanning transmission electron microscopy

DNA repair protein RAD51 homolog 1/DNA Complex

meitotic homologous recombination, DNA repair, ATPase, RECOMBINATION