|Entry||Database: EMDB / ID: 3643|
|Title||Complex of CMG helicase with polymerase epsilon lacking the catalytic domain of Pol2|
|Sample||Complex of CMG helicase with polymerase epsilon where the catalytic domain has been deleted|
|Source||Saccharomyces cerevisiae / Yeast / yeast /|
|Method||single particle reconstruction / 29.6 Å resolution|
|Authors||Zhou JC / Janska A / Goswami P / Renault L / Abid Ali F / Kotecha A / Diffley JFX / Costa A|
|Citation||Journal: Proc. Natl. Acad. Sci. U.S.A. / Year: 2017|
Title: CMG-Pol epsilon dynamics suggests a mechanism for the establishment of leading-strand synthesis in the eukaryotic replisome.
Authors: Jin Chuan Zhou / Agnieszka Janska / Panchali Goswami / Ludovic Renault / Ferdos Abid Ali / Abhay Kotecha / John F X Diffley / Alessandro Costa
Abstract: The replisome unwinds and synthesizes DNA for genome duplication. In eukaryotes, the Cdc45-MCM-GINS (CMG) helicase and the leading-strand polymerase, Pol epsilon, form a stable assembly. The ...The replisome unwinds and synthesizes DNA for genome duplication. In eukaryotes, the Cdc45-MCM-GINS (CMG) helicase and the leading-strand polymerase, Pol epsilon, form a stable assembly. The mechanism for coupling DNA unwinding with synthesis is starting to be elucidated, however the architecture and dynamics of the replication fork remain only partially understood, preventing a molecular understanding of chromosome replication. To address this issue, we conducted a systematic single-particle EM study on multiple permutations of the reconstituted CMG-Pol epsilon assembly. Pol epsilon contains two flexibly tethered lobes. The noncatalytic lobe is anchored to the motor of the helicase, whereas the polymerization domain extends toward the side of the helicase. We observe two alternate configurations of the DNA synthesis domain in the CMG-bound Pol epsilon. We propose that this conformational switch might control DNA template engagement and release, modulating replisome progression.
|Date||Deposition: Mar 20, 2017 / Header (metadata) release: Mar 29, 2017 / Map release: Apr 12, 2017 / Last update: Apr 26, 2017|
|Structure viewer||EM map: |
Downloads & links
|File||emd_3643.map.gz (map file in CCP4 format, 8389 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 3.45 Å|
CCP4 map header:
-Entire Complex of CMG helicase with polymerase epsilon where the catalyt...
|Entire||Name: Complex of CMG helicase with polymerase epsilon where the catalytic domain has been deleted|
Number of components: 1
-Component #1: protein, Complex of CMG helicase with polymerase epsilon where th...
|Specimen||Specimen state: particle|
|Sample solution||pH: 7.6|
|Vitrification||Cryogen name: NONE|
-Electron microscopy imaging
Model: Tecnai Spirit / Image courtesy: FEI Company
|Imaging||Microscope: FEI TECNAI SPIRIT|
|Electron gun||Electron source: TUNGSTEN HAIRPIN / Accelerating voltage: 120 kV / Electron dose: 35 e/Å2 / Illumination mode: SPOT SCAN|
|Lens||Imaging mode: OTHER|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN ULTRASCAN 1000 (2k x 2k)|
|Processing||Method: single particle reconstruction / Number of projections: 15526|
|3D reconstruction||Resolution: 29.6 Å / Resolution method: FSC 0.143 CUT-OFF|
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