2020-06-17 REL PDBe PDBe PDBe 2020-03-05 2020-05-06 2020-05-06 2020-06-17 Medical Research Council (MRC, United Kingdom) MC_UP_1201/12 United Kingdom Conformation2_Mcm2,5-ssDNA_MultiBody Baretic D Jenkyn-Bedford M Baretic D Jenkyn-Bedford M Aria V Cannone G Skehel M Yeeles JTP Cryo-EM Structure of the Fork Protection Complex Bound to CMG at a Replication Fork. Mol.Cell US 78 926 940.e13 2020 32369734 doi:10.1016/j.molcel.2020.04.012 1097-2765 2168 MOCEFL EMD-10730 associated EM volume Conformation 2 of CMG-Csm3-Tof1-Mrc1-Ctf4 with a fork DNA Conformation 2 of CMG-Csm3-Tof1-Mrc1-Ctf4 with a fork DNA 0 1 2 3 4 5 6 7
Mcm2,5 domains with bound fragment of ssDNA
1.4 CMG-Csm3-Tof1-Mrc1-Ctf4 1 1 3 4 5 6 7 Saccharomyces cerevisiae Saccharomyces cerevisiae DNA 1 2 synthetic construct synthetic construct singleParticle particle 0.5 7.5 25.0 HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid 150.0 NaOAc sodium acetate 0.5 TCEP tris(2-carboxyethyl)phosphine 0.1 AMP-PNP adenylyl-imidodiphosphate 0.7 Mg(OAc)2 magnesium acetate 0.005 TWEEN20 Polyoxyethylene (20) sorbitan monolaurate Quantifoil R2/2 COPPER 400 CARBON CONTINUOUS 5.0 PLASMA CLEANING
plasma current of 15 mA
 ETHANE 80 277.15 HOMEMADE PLUNGER
Three microlitres of sample was applied on a grid and incubated for 15-30 s at 4 degC before manually blotting with filter paper for 10 s and plunge-freezing in liquid ethane..
In vitro reconstitution from individual components: CMG, Csm3-Tof1, Mrc1, Ctf4 and a DNA fork
 FEI TITAN KRIOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 300 50.0 2.7 1.4 0.5 2.6 3.0 130000.0 FEI TITAN KRIOS AUTOGRID HOLDER NITROGEN GIF Bioquantum 20 GATAN K2 QUANTUM (4k x 4k) COUNTING 1-20 2 6682 7.0 37.0 1 632000 Gctf 0.50 5U8S 2 C1 FOURIER SPACE 3.7 FSC 0.143 CUT-OFF RELION 3.0 184000 MAXIMUM LIKELIHOOD RELION 3.0 MAXIMUM LIKELIHOOD RELION 3.0 5 RELION 3.0 emd_10730.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 360 360 360 0 0 0 360 360 360 377.64 377.64 377.64 90.0 90.0 90.0 X Y Z -0.030374365 0.061923128 -0.0002571257 0.0016520123 1.049 1.049 1.049 0.01 AUTHOR Conformation2-multibody-Mcm2,5-ssDNA 5U8S 2 5U8S 5 OTHER FSC 0.5 REAL 20