Characterization of a late expression gene, Open reading frame 128 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus.
Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearSNPV) is a single embedded NPV pathogenic to the bollworm, which is a major agricultural pest in many areas around the world. Ha128 homologues have been identified in all completely sequenced lepidopteran NPV's, but no homologue has been found in a granulovirus (GV) and it is thus considered as a lepidopteran NPV-specific gene. In the HearSNPV-C1 genome Ha128 is located between 120,252 and 121,052 bp and encodes a putative protein of 266 amino acid residues with a predicted molecular weight of 30.5 kDa. Ha128 transcripts in HearSNPV-infected HzAM1 cells could be detected from 24 to 120 h post-infection (p.i.) by Northern blot. The Ha128 protein was detected at 24 h p.i. and remained detectable until 120 h p.i. by western blot using an anti-GST-Ha128 antiserum. The expression of Ha128 was inhibited in the presence of Ara-C, an inhibitor of viral DNA synthesis. These results together indicated that Ha128 was a late gene. The product of Ha128 was found to have an Mr of about 31 kDa, in agreement with the predicted molecular weight. Immunoflorescence using anti-GST-Ha128 serum showed that Ha128 was located in cytoplasm. GST pull-down and co-immunoprecipitation showed that at least two potential host proteins interacted with Ha128. In conclusion, Ha128 is a late protein localized in the cytoplasm of infected cells that may interact with host proteins.
An
S-H
SH
Institute of Applied Entomology, Zhejiang University, Hangzhou, P.R. China.
Wang
D
D
Zhang-Nv
Y
Y
Guo
Z-J
ZJ
Xu
H-J
HJ
Sun
J-X
JX
Zhang
C-X
CX
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D017124:Conserved Sequence; D003593:Cytoplasm / Q000737:chemistry; D005814:Genes, Viral; D047468:Immunoprecipitation; D007915:Lepidoptera / Q000821:virology; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D016366:Open Reading Frames; D011485:Protein Binding; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D016415:Sequence Alignment; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins