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Plant Pathology

2017

Tombusvirus

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Co-Opting Atp-Generating Glycolytic Enzyme Pgk1 Phosphoglycerate Kinase Facilitates The Assembly Of Viral Replicase Complexes, K. Reddisiva Prasanth, Chingkai Chuang, Peter D. Nagy Oct 2017

Co-Opting Atp-Generating Glycolytic Enzyme Pgk1 Phosphoglycerate Kinase Facilitates The Assembly Of Viral Replicase Complexes, K. Reddisiva Prasanth, Chingkai Chuang, Peter D. Nagy

Plant Pathology Faculty Publications

The intricate interactions between viruses and hosts include exploitation of host cells for viral replication by using many cellular resources, metabolites and energy. Tomato bushy stunt virus (TBSV), similar to other (+)RNA viruses, induces major changes in infected cells that lead to the formation of large replication compartments consisting of aggregated peroxisomal and ER membranes. Yet, it is not known how TBSV obtains the energy to fuel these energy-consuming processes. In the current work, the authors discovered that TBSV co-opts the glycolytic ATP-generating Pgk1 phosphoglycerate kinase to facilitate the assembly of new viral replicase complexes. The recruitment of Pgk1 into …


The Role Of Co-Opted Escrt Proteins And Lipid Factors In Protection Of Tombusviral Double-Stranded Rna Replication Intermediate Against Reconstituted Rnai In Yeast, Nikolay Kovalev, Jun-Ichi Inaba, Zhenghe Li, Peter D. Nagy Jul 2017

The Role Of Co-Opted Escrt Proteins And Lipid Factors In Protection Of Tombusviral Double-Stranded Rna Replication Intermediate Against Reconstituted Rnai In Yeast, Nikolay Kovalev, Jun-Ichi Inaba, Zhenghe Li, Peter D. Nagy

Plant Pathology Faculty Publications

Reconstituted antiviral defense pathway in surrogate host yeast is used as an intracellular probe to further our understanding of virus-host interactions and the role of co-opted host factors in formation of membrane-bound viral replicase complexes in protection of the viral RNA against ribonucleases. The inhibitory effect of the RNA interference (RNAi) machinery of S. castellii, which only consists of the two-component DCR1 and AGO1 genes, was measured against tomato bushy stunt virus (TBSV) in wild type and mutant yeasts. We show that deletion of the co-opted ESCRT-I (endosomal sorting complexes required for transport I) or ESCRT-III factors makes …


Sterol Binding By The Tombusviral Replication Proteins Is Essential For Replication In Yeast And Plants, Kai Xu, Peter D. Nagy Apr 2017

Sterol Binding By The Tombusviral Replication Proteins Is Essential For Replication In Yeast And Plants, Kai Xu, Peter D. Nagy

Plant Pathology Faculty Publications

Membranous structures derived from various organelles are important for replication of plus-stranded RNA viruses. Although the important roles of co-opted host proteins in RNA virus replication have been appreciated for a decade, the equally important functions of cellular lipids in virus replication have been gaining full attention only recently. Previous work with Tomato bushy stunt tombusvirus (TBSV) in model host yeast has revealed essential roles for phosphatidylethanolamine and sterols in viral replication. To further our understanding of the role of sterols in tombusvirus replication, in this work we showed that the TBSV p33 and p92 replication proteins could bind to …


Three-Dimensional Imaging Of The Intracellular Assembly Of A Functional Viral Rna Replicase Complex, Isabel Fernández De Castro, José J. Fernández, Daniel Barajas, Peter D. Nagy, Cristina Risco Jan 2017

Three-Dimensional Imaging Of The Intracellular Assembly Of A Functional Viral Rna Replicase Complex, Isabel Fernández De Castro, José J. Fernández, Daniel Barajas, Peter D. Nagy, Cristina Risco

Plant Pathology Faculty Publications

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 …