Bombyx mori nucleopolyhedrovirus orf8 encodes a nucleic acid binding protein that colocalizes with IE1 during infection.
This report describes the characterization of the Bombyx mori nucleopolyhedrovirus (BmNPV) orf8 gene. Immunoblot analyses demonstrated that orf8 was expressed as an early gene. The ORF8 protein accumulated in the nucleus, and was maintained at relatively constant levels from 4 to 24 h postinfection. Immunoblot analysis failed to detect ORF8 protein associated with budded virus and occlusion derived virus. In addition, immunohistochemical analysis by confocal microscopy showed that ORF8 protein colocalized with IE1 to specific nuclear foci throughout infection. To further examine the function of ORF8, a reporter gene was inserted into the orf8 reading frame. One orf8 disruption mutant (BmD8), which expressed the N-terminal half of ORF8, was isolated. However, it was not possible to isolate a null mutant, suggesting that orf8 may have an important role during viral infection. Single-step growth curves showed that BV production was reduced in BmD8 infected cells. Biochemical analyses indicated that ORF8 bound to nucleic acids. Together, these results suggest that BmNPV ORF8 may be involved in viral DNA replication and/or transcription.
Imai
N
N
Laboratory of Molecular Entomology and Baculovirology, RIKEN, Hirosawa, Japan.
Kurihara
M
M
Matsumoto
S
S
Kang
W-K
WK
D000818:Animals; D012831:Bombyx / Q000821:virology*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000378:metabolism; D016366:Open Reading Frames; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
C069183:Ac-IE-1 protein, Autographa californica nucleopolyhedrovirus; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D015534:Trans-Activators