Qualitative proteomic analysis of Tipula oleracea nudivirus occlusion bodies
Résumé
Nudiviruses are arthropod-specific large double-stranded circular DNA viruses, related to baculoviruses, which replicate in the nucleus of the cells they infect. Up to date six fully sequenced nudiviral genomes are available in databases and protein profile from nudivirus particles were mainly characterized by polyacrylamide gel electrophoresis. However, only few direct matches were completed between genomic and proteomic data to the exception of the major occlusion body protein from Penaeus monodon nudivirus (PmNV) and four nucleocapsid proteins from Helicoverpa zea nudivirus 2 (HzNV-2). Function of predicted nudiviral proteins is still inferred from what is known from baculoviruses or endogenous nudiviruses (i.e. bracoviruses). Tipula oleracea nudivirus (ToNV) is the causative agent of crane fly nucleopolyhedrosis. With PmNV, ToNV is the second fully sequenced nudivirus to be described as forming occlusion bodies. Protein profile revealed by Coomassie-stained SDS-PAGE is quite similar to those observed for other nudiviruses with five major protein bands of about 75, 48, 35, 25 and 12 kDa. Proteomic analysis using on-line nanoflow liquid chromatography tandem high resolution mass spectrometry revealed ToNV occlusion bodies are composed of 52 viral proteins, the most abundant of which are the functional homolog of baculovirus polyhedrin/granulin and the homologs of three HzNV-2 predicted proteins: the two virion structural proteins 34K (Hz2V052, the baculovirus capsid protein VP39 homolog) and 11K (Hz2V025); and the hypothetical protein Hz2V079, a newly identified nudivirus core gene product.
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