Effects of substituting granulin or a granulin-polyhedrin chimera for polyhedrin on virion occlusion and polyhedral morphology in Autographa californica multinucleocapsid nuclear polyhedrosis virus.
Substitution of granulin from the Trichoplusia ni granulosis virus (TnGV) for polyhedrin of the Autographa californica multinucleocapsid nuclear polyhedrosis virus (AcMNPV) yielded a few very large (2 to 5 micron) cuboidal inclusions in the cytoplasm and nucleus of infected cells. These polyhedra lacked the beveled edges characteristic of wild-type AcMNPV polyhedra, contained fractures, and occluded few virions. Placing a nuclear localization signal (KRKK) in granulin directed more granulin to the nucleus and resulted in more structurally uniform cuboidal inclusions in which no virions were observed. A granulin-polyhedrin chimera produced tetrahedral occlusions with more virions than granulin inclusions but many fewer than wild-type polyhedra. Despite the unusual structure of the granulin and granulin-polyhedrin inclusions, they interacted with AcMNPV p10 fibrillar structures and electron-dense spacers that are precursors of the polyhedral calyx. The change in inclusion shape obtained with the granulin-polyhedrin chimera demonstrates that the primary amino acid sequence affects occlusion body shape, but the large cuboidal inclusions formed by granulin indicate that the amino acid sequence is not the only determinant. The failure of granulin or the granulin-polyhedrin chimera to properly occlude AcMNPV virions suggests that specific interactions occur between polyhedrin and other viral proteins which facilitate normal virion occlusion and occlusion body assembly and shape in baculoviruses.
Eason
J E
JE
Interdepartmental Graduate Program in Genetics, University of California, Riverside, California 92521, USA.
Hice
R H
RH
Johnson
J J
JJ
Federici
B A
BA
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000502:physiology* / Q000648:ultrastructure; D000077183:Occlusion Body Matrix Proteins; D016366:Open Reading Frames; D011993:Recombinant Fusion Proteins; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D015678:Viral Structural Proteins; D014771:Virion / Q000502:physiology*; D019065:Virus Assembly
D000077183:Occlusion Body Matrix Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C000625703:granulin protein, granulovirus; C009829:polyhedrin protein, Nucleopolyhedrovirus