Journal article
Localized reconstruction of subunits from electron cryomicroscopy images of macromolecular complexes
- 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.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 1.4MB, Terms of use)
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- Publisher copy:
- 10.1038/ncomms9843
Authors
+ European Research Council (ERC) under the European Union’s Horizon 2020 research
and innovation programme
More from this funder
- Funding agency for:
- Huiskonen, J
- Grant:
- 649053
- Publisher:
- Nature Publishing Group
- Journal:
- Nature Communications More from this journal
- Volume:
- 6
- Article number:
- 8843
- Publication date:
- 2015-11-04
- Acceptance date:
- 2015-10-08
- DOI:
- EISSN:
-
2041-1723
- ISSN:
-
2041-1723
- Language:
-
English
- Keywords:
- Pubs id:
-
pubs:575131
- UUID:
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uuid:f577caf5-f8b6-49c4-83ad-e2ddc3e3ecab
- Local pid:
-
pubs:575131
- Source identifiers:
-
575131
- Deposit date:
-
2016-04-22
Terms of use
- Copyright holder:
- Macmillan Publishers Limited
- Copyright date:
- 2015
- Notes:
- © 2015 Macmillan Publishers Limited. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
- Licence:
- CC Attribution (CC BY)
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