The Nedd4-Type Rsp5p Ubiquitin Ligase Inhibits Tombusvirus Replication by Regulating Degradation of the p92 Replication Protein and Decreasing the Activity of the Tombusvirus Replicase
- 15 November 2009
- journal article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 83 (22), 11751-11764
- https://doi.org/10.1128/jvi.00789-09
Abstract
Recent in vitro proteomics screens revealed that many host proteins could interact with the replication proteins of Tomato bushy stunt virus (TBSV), which is a small, plus-stranded RNA virus (Z. Li, D. Barajas, T. Panavas, D. A. Herbst, and P. D. Nagy, J. Virol. 82:6911-6926, 2008). To further our understanding of the roles of host factors in TBSV replication, we have tested the effect of Rsp5p, which is a member of the Nedd4 family of E3 ubiquitin ligases. The full-length Rsp5p, via its WW domain, is shown to interact with p33 and the central portion of p92 pol replication proteins. We find that overexpression of Rsp5p inhibits TBSV replication in Saccharomyces cerevisiae yeast, while downregulation of Rsp5p leads to increased TBSV accumulation. The inhibition is caused by Rsp5p-guided degradation of p92 pol , while the negative effect on the p33 level is less pronounced. Interestingly, recombinant Rsp5p also inhibits TBSV RNA replication in a cell-free replication assay, likely due to its ability to bind to p33 and p92 pol . We show that the WW domain of Rsp5p, which is involved in protein interactions, is responsible for inhibition of TBSV replication, whereas the HECT domain, involved in protein ubiquitination, is not necessary for Rsp5p-mediated inhibition of viral replication. Overall, our data suggest that direct binding between Rsp5p and p92 pol reduces the stability of p92 pol , with consequent inhibition of TBSV replicase activity.Keywords
This publication has 45 references indexed in Scilit:
- A Key Role for Heat Shock Protein 70 in the Localization and Insertion of Tombusvirus Replication Proteins to Intracellular MembranesJournal of Virology, 2009
- A Functional Genomic Screen Identifies Cellular Cofactors of Hepatitis C Virus ReplicationCell Host & Microbe, 2009
- Translation elongation factor 1A is a component of the tombusvirus replicase complex and affects the stability of the p33 replication co-factorVirology, 2009
- In vitro assembly of the Tomato bushy stunt virus replicase requires the host Heat shock protein 70Proceedings of the National Academy of Sciences, 2008
- Cdc34p Ubiquitin-Conjugating Enzyme Is a Component of the Tombusvirus Replicase Complex and Ubiquitinates p33 Replication ProteinJournal of Virology, 2008
- Drosophila RNAi screen identifies host genes important for influenza virus replicationNature, 2008
- Authentic Replication and Recombination of Tomato Bushy Stunt Virus RNA in a Cell-Free Extract from YeastJournal of Virology, 2008
- Regulation of catalytic activities of HECT ubiquitin ligasesBiochemical and Biophysical Research Communications, 2007
- Identification of Essential Host Factors Affecting Tombusvirus RNA Replication Based on the Yeast Tet Promoters Hughes CollectionJournal of Virology, 2006
- THE ESCRT COMPLEXES: Structure and Mechanism of a Membrane-Trafficking NetworkAnnual Review of Biophysics and Biophysical Chemistry, 2006