RNA-binding proteins that inhibit RNA virus infection.
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.
Zhu
Jian
J
Department of Pharmacology and Molecular Sciences and High Throughput Biology Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Gopinath
Kodetham
K
Murali
Ayaluru
A
Yi
Guanghui
G
Hayward
S Diane
SD
Zhu
Heng
H
Kao
Cheng
C
D020816:Amino Acid Motifs / Q000235:genetics; D015152:Blotting, Northern; D017795:Bromovirus / Q000235:genetics* / Q000502:physiology; D019751:Intramolecular Transferases / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D040081:Protein Array Analysis; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D014026:Nicotiana / Q000821:virology; D019065:Virus Assembly / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects*
D008840:Microfilament Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; C505432:APP1 protein, S cerevisiae; D019751:Intramolecular Transferases; C023199:tRNA-pseudouridine synthase I