Structural homology among four nodaviruses as deduced by sequencing and X-ray crystallography.
The genomic RNA2s of nodaviruses encode a single gene, that of protein alpha, the precursor of virion proteins beta and gamma. We compared the sequences of the RNA2s of the nodaviruses, black beetle virus (BBV), flock house virus, boolarra virus and nodamura virus, with the objective of identifying homologies in the primary and secondary structure of these RNAs and in the structure of their encoded protein. The sequences of the four RNAs were found to be similar, so that homologous regions relating to translation and RNA replication were readily identified. However, the overall, secondary structures in solution, deduced from calculations of optimal Watson-Crick base-pairing configurations, were very different for the four RNAs. We conclude that a particular, overall, secondary structure in solution within host cells is not required for virus viability. The partially refined X-ray structure of BBV (R = 26.4% for the current model) was used as a framework for comparing the structure of the encoded proteins of the four viruses. Mapping of the four protein sequences onto the BBV capsid showed many amino acid differences on the outer surface, indicating that the exteriors of the four virions are substantially different. Mapping in the beta-barrel region showed an intermediate level of differences, indicating that some freedom in choice of amino acid residues is possible there although the basic framework of the capsids is evidently conserved. Mapping onto the interior surface of the BBV capsid showed a high degree of conservation of amino acid residues, particularly near the protein cleavage site, implying that that region is nearly identical in all four virions and has an essential role in virion maturation, and also suggests that all four capsid interior surfaces have similar surfaces exposed to the viral RNA. Apart from a small portion of the C promoter, the amino terminus of the BBV protein (residues 1 to 60) is crystallographically disordered and the amino acid residues in that region are not well conserved. The disordered portion of the BBV protein clearly projects from the capsid inner surface into the interior of the virion, the region occupied by the viral RNA. In all four viruses, residues 1 to 60 had a high proportion of basic residues, suggesting a virus-specific interaction of the amino terminus with the virion RNA.
Kaesberg
P
P
Institute for Molecular Virology, University of Wisconsin, Madison 53706.
Dasgupta
R
R
Sgro
J Y
JY
Wery
J P
JP
Selling
B H
BH
Hosur
M V
MV
Johnson
J E
JE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002213:Capsid; D002478:Cells, Cultured; D004331:Drosophila melanogaster; D005814:Genes, Viral; D007304:Insect Viruses / Q000032:analysis / Q000235:genetics*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D012367:RNA, Viral / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014764:Viral Proteins; D014780:Viruses / Q000032:analysis / Q000235:genetics*; D014961:X-Ray Diffraction
D012367:RNA, Viral; D014764:Viral Proteins