Characterization of the 25K FP gene of the baculovirus Bombyx mori nucleopolyhedrovirus: implications for post-mortem host degradation.
Mutagenesis experiments on the baculovirus Bombyx mori nucleopolyhedrovirus (BmNPV) using 5-bromo-2'-deoxyuridine generated five mutants with a 'few polyhedra' (FP) phenotype. Sequence analysis of the 25K gene homologue of the BmNPV FP mutants revealed nucleotide substitutions in the coding region. Rescue experiments indicated that the FP phenotype of the BmNPV mutants resulted from mutations in the 25K coding region. Effects of infection by these FP mutants were analysed following injection of the viruses into silkworm (B. mori) larvae. Compared to infection with wild-type virus, infection with each FP mutant resulted in reduced host degradation (liquefaction). The degree to which liquefaction was blocked corresponded to the degree of functional disruption of the 25K gene product and to the extent to which polyhedron production was reduced. Electron microscopy revealed that (1) polyhedron production was reduced, (2) very few virions were occluded and those that were lacked envelopes, and (3) the basal lamina of fat-body tissue was not destroyed by infection and accumulations of virions occurred along the membrane. Typical NPV-induced liquefaction was observed following infection with a polyhedrin deletion mutant, indicating that host degradation was not related to polyhedron production. These results suggest that (1) the 25K gene product is involved in the host degradation process caused by virus infection and (2) the FP phenotype is an indirect result of disruption of the 25K gene; activation or suppression of a specific host or viral gene related to tissue degradation and polyhedron formation may be involved.
Katsuma
S
S
Noguchi
Y
Y
Zhou
C L
CL
Kobayashi
M
M
Maeda
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000196:embryology / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology*; D001973:Bromodeoxyuridine / Q000378:metabolism; D002460:Cell Line; D017353:Gene Deletion; D005814:Genes, Viral / Q000235:genetics / Q000502:physiology*; D005816:Genetic Complementation Test; D007814:Larva / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D008854:Microscopy, Electron; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure; D016366:Open Reading Frames / Q000235:genetics; D010641:Phenotype; D017422:Sequence Analysis, DNA; D014162:Transfection; D019065:Virus Assembly / Q000235:genetics
D001973:Bromodeoxyuridine