Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 104, Issue 9, Pages 3129-3134Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0611617104
Keywords
brome mosaic virus; protein-RNA interaction; pseudouricline synthase 4; yeast proteome chip; viral RNA replication
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Funding
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [0925401] Funding Source: National Science Foundation
- NCRR NIH HHS [U54RR020839-01, U54 RR020839] Funding Source: Medline
- NIAID NIH HHS [R21 AI070740, 1R21AI070740-01] Funding Source: Medline
- NIGMS NIH HHS [1R01GM076102-01A1, R01 GM076102] Funding Source: Medline
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Arrays of > 5,000 Saccharomyces cerevisiae proteins were screened to identify proteins that can preferentially bind a small RNA hairpin that contains a clamped adenine motif (CAM). A CAM is required for the replication of Brome Mosaic Virus (BMV), a plant-infecting RNA virus that can replicate in S. cerevisiae. Several hits were selected for further characterization in Nicotiana benthamiana. Pseudouridine Synthase 4 (Pus4) and the Actin Patch Protein 1 (App1) modestly reduced BMV genomic plus-strand RNA accumulation, but dramatically inhibited BMV systemic spread in plants. Pus4 also prevented the encapsidation of a BMV RNA in plants and the reassembly of BMV virions in vitro. These results demonstrate the feasibility of using proteome arrays to identify specific RNA-binding proteins for antiviral activities. Furthermore, the effects of Pus4 suggest that the CAM-containing RNA motif provides a regulatory link between RNA replication and encapsidation.
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