期刊
JOURNAL OF CELL SCIENCE
卷 130, 期 1, 页码 260-268出版社
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/jcs.181586
关键词
3D electron microscopy; Tomography; Metal-tagging transmission electron microscopy; Clonable tag; Viral replication factory
类别
资金
- Spanish Ministry of Economy and Competitiveness [BIO2012-33314, TIN2012-37483]
- US National Science Foundation [MCB-1122039]
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [1517751] Funding Source: National Science Foundation
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.
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