1. Screening a yeast library of temperature-sensitive mutants reveals a role for actin in tombusvirus RNA recombination
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K. Reddisiva Prasanth, Peter D. Nagy, Jannine Baker, Isabel Fernández de Castro Martín, and Nikolay Kovalev
- Subjects
0301 basic medicine ,Tombusvirus ,Saccharomyces cerevisiae Proteins ,FLP-FRT recombination ,Saccharomyces cerevisiae ,RNA-dependent RNA polymerase ,Virus Replication ,Genetic recombination ,03 medical and health sciences ,Virology ,Genetics ,Recombination, Genetic ,biology ,Temperature ,Helicase ,RNA ,biology.organism_classification ,Actins ,030104 developmental biology ,Viral replication ,Actin-Related Protein 3 ,Host-Pathogen Interactions ,Mutation ,biology.protein - Abstract
Genetic recombination in RNA viruses drives the evolutionary arms race with host׳s antiviral strategies and recombination also facilitates adaptation of viruses to new hosts. In this paper, the authors used tombusvirus and a temperature-sensitive (ts) mutant library of yeast to identify 40 host proteins affecting viral recombination in yeast model host. Subsequent detailed analysis with two identified actin-related proteins, Act1p and Arp3p, has revealed that the wt actin network helps TBSV to maintain low level viral recombination. Pharmacological inhibition of actin in plant protoplasts confirmed the role of the actin network in tombusvirus recombination. An in vitro approach revealed the altered activity of the tombusvirus replicase in the presence of mutated Act1p. The authors show more efficient recruitment of a cellular DEAD-box helicase, which enhances tombusvirus recombination, into the membrane-bound replicase in Act1p mutant yeast. Overall, this work shows that the actin network affects tombusvirus recombination in yeast and plant cells.
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