Identification and sequencing of the spheroidin gene of Choristoneura biennis entomopoxvirus.
Entomopoxviruses are a class of insect viruses whose virions are embedded in cytoplasmic occlusion bodies. The major component of these protective complexes is a protein called spheroidin. An open reading frame encoding the spheroidin gene of Choristoneura biennis entomopoxvirus has been identified and sequenced in our laboratory. This protein coding region is 1023 nucleotides long and specifies a polypeptide of 38,500 Da. Spheroidin was purified by SDS polyacrylamide gel electrophoresis, electroeluted, and its amino terminus sequence was determined on a gas phase sequencer. We observed that the first 20 N-terminal amino acids were absent in the mature processed form of the spheroidin molecule. Examination of these 20 residues revealed their hydrophobic nature and close resemblance to the consensus signal peptide sequence which is commonly found on membrane proteins. The DNA sequence of the spheroidin gene predicted a processed polypeptide with a molecular weight of 36 kDa. However, spheroidin was observed to aggregate in complexes composed of 50-kDa monomers. Intermolecular disulfide bonds were shown to play major roles in the formation and structure of these viral occlusion bodies. The difference in molecular weight between the predicted protein and its counterpart in infected cells is likely due to post-translational modifications. Indeed, two potential asparagine-linked glycosylation sites are present on the spheroidin molecule. The 5' flanking regions of the spheroidin gene and the vaccinia major core protein precursor gene P4b were shown to share substantial homology.
Yuen
L
L
Genetic Engineering Section, National Research Council of Canada, Montreal, Quebec.
Dionne
J
J
Arif
B
B
Richardson
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics*; D004175:Diptera; D004591:Electrophoresis, Polyacrylamide Gel; D007304:Insect Viruses / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011212:Poxviridae / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics; D021382:Protein Sorting Signals / Q000032:analysis / Q000235:genetics; D011506:Proteins / Q000032:analysis / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D012984:Software; D014616:Vaccinia virus / Q000235:genetics; D014758:Viral Core Proteins / Q000235:genetics; D014764:Viral Proteins / Q000032:analysis / Q000235:genetics*; D015678:Viral Structural Proteins
D004279:DNA, Viral; D011498:Protein Precursors; D021382:Protein Sorting Signals; D011506:Proteins; D014758:Viral Core Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C063744:spheroidin protein, Entomopoxvirus