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Localized reconstruction of subunits from electron cryomicroscopy images of macromolecular complexes

DOI: 10.1038/ncomms9843 DOI Help
PMID: 26534841 PMID Help

Authors: Serban L Ilca (University of Oxford) , Abhay Kotecha (University of Oxford) , Xiaoyu Sun (University of Helsinki) , Minna M. Poranen (University of Helsinki) , David L Stuart (Diamond Light Source) , Juha T Huiskonen (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 6

State: Published (Approved)
Published: November 2015

Open Access Open Access

Abstract: Electron cryomicroscopy can yield near-atomic resolution structures of highly ordered macromolecular complexes. Often however some subunits bind in a flexible manner, have different symmetry from the rest of the complex, or are present in sub-stoichiometric amounts, limiting the attainable resolution. Here we report a general method for the localized three-dimensional reconstruction of such subunits. After determining the particle orientations, local areas corresponding to the subunits can be extracted and treated as single particles. We demonstrate the method using three examples including a flexible assembly and complexes harbouring subunits with either partial occupancy or mismatched symmetry. Most notably, the method allows accurate fitting of the monomeric RNA-dependent RNA polymerase bound at the threefold axis of symmetry inside a viral capsid, revealing for the first time its exact orientation and interactions with the capsid proteins. Localized reconstruction is expected to provide novel biological insights in a range of challenging biological systems.

Subject Areas: Biology and Bio-materials

Technical Areas: