Analysis of in vitro Translation of Bean Yellow Mosaic Virus RNA: Inhibition of Proteolytic Processing by Antiserum to the 49K Nuclear Inclusion Protein
In vitro translation of bean yellow mosaic virus (BYMV) RNA in rabbit reticulocyte lysate without dithiothreitol (DTT) resulted in the accumulation of high Mr products. These were readily processed to low Mr mature proteins after the addition of DTT followed by a 2 h incubation. Immunoprecipitation analyses of the partially processed high Mr products with antisera to the 49K and 54K nuclear inclusion (NI) proteins, cylindrical inclusion (CI) protein, tobacco vein mottling virus helper component (HC) protein and capsid protein (CP) resolved the following BYMV RNA genome map: 5′ end, 32K unidentified protein, 48K HC, 42K unidentified protein, 73K CI, 49K NI, 54K NI, 32K CP, 3′ end. The addition of the 49K NI protein antiserum inhibited the proteolytic processing of the high Mr translation products. The inhibition of processing was diminished by diluting the 49K NI antiserum or by adding purified 49K NI protein. Preimmune serum or the antisera to the 54K NI, CI and capsid proteins were not as effective in inhibiting the proteolytic processing. These results provide evidence that the 49K NI protein or a related protein may have a protease function in the processing of the potyvirus polyproteins.
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Analysis of in vitro Translation of Bean Yellow Mosaic Virus RNA: Inhibition of Proteolytic Processing by Antiserum to the 49K Nuclear Inclusion Protein