Streaming video does not reproduce the original video quality. Download the video using the link below to view at original quality.The video illustrates the transition from free monomeric Pol I to elongating Pol I. The four major polymerase modules core, jaw-lobe, clamp and shelf are colored in grey, blue, yellow and pink, respectively. The movement of the rigid clamp-shelf module is highlighted together with the refolding of the bridge helix (green). The adjacent dimer of model of the crystal structure (PDB 4C2M) is shown briefly before the focus is set on the domain movements. Furthermore, the expander (light green), downstream DNA (blue), the DNA-RNA hybrid (blue-red) and the C-terminal domain of A12.2 are visualized.
More videos from this article
These authors contributed equally to this work.
- Simon Neyer &
- Michael Kunz
These authors jointly supervised this work.
- Patrick Cramer &
- Achilleas S. Frangakis
Max-Planck-Institute for Biophysical Chemistry, Department of Molecular Biology, Am Fassberg 11, 37077 Göttingen, Germany
- Simon Neyer,
- Merle Hantsche,
- Christoph Engel &
- Patrick Cramer
Buchmann Institute for Molecular Life Sciences and Institute for Biophysics, Goethe University Frankfurt, Max-von-Laue Str. 15, Frankfurt 60438, Germany
- Michael Kunz,
- Christian Geiss,
- Victor-Valentin Hodirnau,
- Anja Seybert,
- Margot P. Scheffer &
- Achilleas S. Frangakis
S.N. planned and carried out the single particle sample preparation, data collection and data analysis. M.K. planned and carried out the tomographic data analysis. C.G. carried out the sample preparation for tomography. M.H. advised on structure determination procedures. V.V.H. advised on and carried out sample preparation for tomography. A.S. advised on sample preparation for tomography. C.E. advised on biochemical procedures. M.P.S. advised on tomographic data analysis. P.C. designed and supervised research, and supervised single particle structure determination. A.S.F. designed and supervised research, supervised single particle data collection and performed tomographic data collection and analysis. S.N., P.C. and A.S.F. prepared the manuscript, with contributions from all authors.
Competing financial interests
The authors declare no competing financial interests.
Reviewer Information: Nature thanks R. Ebright, E. Nogales and E. Nudler for their contribution to the peer review of this work.
Extended Data Figure 1: Preparation of Pol I elongation complex (EC) for single-particle cryo-EM.Hover over figure to zoom
Extended Data Figure 2: Single-particle cryo-EM particle sorting pipeline.Hover over figure to zoom
Extended Data Figure 3: Quality of single-particle cryo-EM reconstructions.Hover over figure to zoom
Extended Data Figure 4: Comparison of all eukaryotic and the bacterial elongation complexes.Hover over figure to zoom
Extended Data Figure 5: Additional details on Pol I EC.Hover over figure to zoom
Extended Data Figure 6: Free monomeric Pol I single-particle cryo-EM structure.Hover over figure to zoom
Extended Data Figure 7: Yeast cells, lysed to leak their nucleoplasm, prepared with negative stain and visualized under cryo conditions.Hover over figure to zoom
Extended Data Figure 8: Relative positions of polymerases towards each other and of protruding nucleic acids.Hover over figure to zoom
Extended Data Figure 9: Comparisons between cryo-electron tomography and single-particle cryo-EM structures.Hover over figure to zoom
Video 1: Conformational changes between free Pol I and Pol I EC
Extended Data Table 1: Model refinement statisticsHover over figure to zoom