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. 2020 Jun 25;48(14):8128–8145. doi: 10.1093/nar/gkaa541

Figure 1.

Figure 1.

Cryo-EM structure of the Pol2CAT/Ctf18-1-8 complex. (A) Domain architecture and molecular context of Pol2 and Ctf18-RFC. CMGE refers to the Cdc45-Mcm2-7-GINS helicase in complex with the non-catalytic subunits of Pol ϵ. TBD – triple barrel domain, WHH – winged helix hook, NTD – N-terminal domain, F – fingers subdomain, C – C-terminal domain. (B) The left panel shows a sharpened cryo-EM map of the complex colored by Ctf18-1-8 subunit as in panel A and Pol2CAT subdomain, with Pol2NTD in magenta, Pol2PALM in gray, Pol2EXO in salmon, Pol2FINGERS in yellow and Pol2THUMB in brown. The right panel shows the refined model of the complex in cartoon representation with the iron-sulfur cluster as spheres. (C) Superposition by Pol2NTD–Pol2EXO of the cryo-EM structure (colored as in panel B) and the Pol2(1-528)/Ctf18-1-8 crystal structure (pdb: 5oki) shown entirely in red. (D) ITC experiment showing titration of 170 μM Ctf18-1-8 into 25 μM Pol2CAT in the presence of 50 μM P1P2. (E) The three interaction patches are shown on the opened surface of the complex as yellow dashed areas. The electrostatic surface was calculated using APBS (80) and shown as a gradient from –4 kT/e (red) to +4 kT/e (blue).