Effects of deletions in the N-terminal basic arm of brome mosaic virus coat protein on RNA packaging and systemic infection.
The first 25 amino acids of brome mosaic virus (BMV) coat protein include 8 basic and no acidic residues and are implicated in binding the encapsidated RNA. Using infectious transcripts from BMV RNA3 cDNA clones, we modified this region of the coat gene. A coat protein mutant with the first 25 amino acids deleted failed to direct either packaging of viral RNA in protoplasts or systemic infection of whole barley plants. Neither symptoms, virions, nor viral RNA was detectable in plants inoculated with this mutant or a mutant with a frameshift mutation in the coat gene. Mutants with the normal start codon changed to AAG or with the first eight codons deleted allowed translation to start at a downstream AUG, resulting in a deletion of the first 7 amino acids of the mature wild-type coat protein. These mutants not only packaged viral RNA in protoplasts but directed symptomatic, systemic infections that developed with normal speed and degree of spread within the host. The AUG-to-AAG point substitution did not revert to the wild type after long-term culture in planta. Wild-type BMV virions were also found to contain small amounts of a protein that coelectrophoresed with the truncated coat protein produced by the viable AAG and eight-codon-deletion mutants. This minor coat protein species presumably arose by infrequent translation initiation at the second AUG in the wild-type coat protein gene. Absence of encapsidation-competent coat protein appeared to stimulate production of nonstructural proteins in protoplast infections.
Sacher
R
R
Institute for Molecular Virology, University of Wisconsin-Madison 53706.
Ahlquist
P
P
D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D002872:Chromosome Deletion; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics* / Q000472:pathogenicity; D009154:Mutation; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics*; D014771:Virion / Q000235:genetics
D012367:RNA, Viral; D015678:Viral Structural Proteins