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J Cell Sci. 2017 Jan 1;130(1):260-268. doi: 10.1242/jcs.181586. Epub 2016 Mar 29.

Three-dimensional imaging of the intracellular assembly of a functional viral RNA replicase complex.

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Cell Structure Laboratory, Centro Nacional de Biotecnología (CNB-CSIC), Campus de Cantoblanco, Madrid 28049, Spain
Department of Structure of Macromolecules, Centro Nacional de Biotecnología (CNB-CSIC), Campus de Cantoblanco, Madrid 28049, Spain.
Department of Plant Pathology, University of Kentucky, Lexington, Kentucky, KY 40546, USA.


Positive-strand RNA viruses, which can be devastating pathogens in humans, animals and plants, replicate their genomes on intracellular membranes. Here, we describe the three-dimensional ultrastructural organization of a tombusvirus replicase in yeast, a valuable model for exploring virus-host interactions. We visualized the intracellular distribution of a viral replicase protein using metal-tagging transmission electron microscopy, a highly sensitive nanotechnology whose full potential remains to be developed. These three-dimensional images show how viral replicase molecules are organized when they are incorporated into the active domains of the intracellular replication compartment. Our approach provides a means to study protein activation mechanisms in cells and to identify targets for new antiviral compounds.


3D electron microscopy; Clonable tag; Metal-tagging transmission electron microscopy; Tomography; Viral replication factory

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