NUCLEOPOLYHEDROVIRUS PATHOLOGY IN SPRUCE BUDWORM LARVAE

2000 ◽  
Vol 132 (5) ◽  
pp. 581-590 ◽  
Author(s):  
J.W. Barrett ◽  
T.R. Ladd ◽  
M.J. Primavera ◽  
A. Retnakaran ◽  
S.S. Sohi ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Fat Body ◽  
Choristoneura Fumiferana ◽  
Spruce Budworm ◽  
Temporal And Spatial Distribution ◽  
Tracheal System ◽  
Tracheal Epithelial Cells ◽  
Multiple Nucleopolyhedrovirus ◽  
Temporal And Spatial ◽  
Green Fluorescent

AbstractChoristoneura fumiferana (Clemens) (Lepidoptera: Tortricidae) multiple nucleopolyhedrovirus (CfMNPV) expressing green fluorescent protein was used to study aspects of nucleopolyhedrovirus infection in the spruce budworm. The temporal and spatial distribution of fluorescence indicated that the virus infected the midgut, entered the tracheal system, and traveled to the epidermis, fat body, and muscles. In contrast to Autographa californica (Speyer) (Lepidoptera: Noctuidae) multiple nucleopolyhedrovirus (AcMNPV) infection, hemocytes from infected C. fumiferana did not exhibit fluorescence until after CfMNPV had passed from the midgut into the tracheae. Therefore the role of hemocytes may be limited during CfMNPV infection. Also the fluorescence pattern spread from the tracheolar cells to tracheal epithelial cells throughout the tracheal system. Our results indicate that the temporal and spatial events involved in CfMNPV infection of C. fumiferana larvae are consistent with those observed in other lepidopteran hosts infected with AcMNPV Minor deviations between these two systems may be attributed to differences in virulence, infection rate, and possibly host range of the virus.

scholarly journals Autographa californica multiple nucleopolyhedrovirus and Choristoneura fumiferana multiple nucleopolyhedrovirus v-cath genes are expressed as pre-proenzymes

2009 ◽  
Vol 90 (4) ◽  
pp. 995-1000 ◽  
Author(s):  
Jeffrey J. Hodgson ◽  
Basil M. Arif ◽  
Peter J. Krell
Keyword(s):  
Fluorescent Protein ◽  
Choristoneura Fumiferana ◽  
Immunoblot Analysis ◽  
Autographa Californica ◽  
Proteolytic Processing ◽  
Post Translational Modifications ◽  
Peptide Cleavage ◽  
Multiple Nucleopolyhedrovirus ◽  
Group I ◽  
Green Fluorescent

Intracellular processing and trafficking of the baculovirus v-cath expressed cathepsin (V-CATH), which lacks canonical targeting signals, are poorly understood. The cathepsins of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), Choristoneura fumiferana multiple nucleopolyhedrovirus (CfMNPV) and most other alphabaculovirus group I nucleopolyhedroviruses have well-conserved N-termini containing overlapping chymotrypsin-cleavage (Y11) and myristoylation (G12) motifs, which are suggestive of proteolytic signal-peptide cleavage to generate proV-CATH and subsequent acylation. To determine proteolytic N-terminal processing of V-CATH, haemagglutinin epitope-coding tags were fused to the 5′ and/or 3′ ends of AcMNPV and CfMNPV v-cath. Immunoblot analysis suggested that a small N-terminal peptide is cleaved for both viruses, indicating that v-cath is expressed as a pre-proenzyme. The two viral homologues undergo similar proteolytic processing, but have different glycosylation or other post-translational modifications. An AcMNPV V-CATH–DsRED fusion protein co-localized to the endoplasmic reticulum with an HDEL motif-containing green fluorescent protein. Based on these findings, pre-proV-CATH processing and trafficking mechanisms are postulated.

Use of Green Fluorescent Protein (GFT)-Chimeras to Study Cytoskeletal Assembly and Dynamics

1998 ◽  
Vol 4 (S2) ◽  
pp. 1010-1011
Author(s):  
G. G. Gundersen ◽  
A. Mikhailov ◽  
J. L. Martys ◽  
L. Ho ◽  
R. K. H. Liem ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Cell Lines ◽  
Intermediate Filaments ◽  
Fluorescent Protein ◽  
Cell Structure ◽  
Β1 Integrin ◽  
Spatial Regulation ◽  
Temporal And Spatial ◽  
Green Fluorescent ◽  
3T3 Cell Lines

The cytoskeleton plays an important role in cell structure, polarity, locomotion and division. Individual elements of the cytoskeleton are composed of subunit proteins which assemble and disassemble in specific places and times within the cell. Knowledge of the temporal and spatial regulation of subunit assembly and disassembly is essential to understanding how the cytoskeleton contributes to cellular activities. The assembly and dynamics of two cytoskeletal structures, namely adhesion plaques (APs) and intermediate filaments (IFs), have been difficult to study by traditional methods. We have generated GFP-chimeras to label these structures and to study their dynamics in motile fibroblasts.To study the dynamics of APs, we prepared stable 3T3 cell lines expressing a GFP-β1 integrin chimera. The chimera was prepared by fusing GFP to the transmembrane and cytoplasmic portions of β1 intergrin, since previous studies had shown that the cytoplasmic tail of β integrins is sufficient to direct integrins to APs.

scholarly journals Autographa californica multiple nucleopolyhedrovirus ORF 23 null mutant produces occlusion-derived virions with fewer nucleocapsids

2009 ◽  
Vol 90 (6) ◽  
pp. 1499-1504 ◽  
Author(s):  
Ian-Ling Yu ◽  
Doug Bray ◽  
Ying-Chu Lin ◽  
Oliver Lung
Keyword(s):  
Fluorescent Protein ◽  
Autographa Californica ◽  
Dapi Staining ◽  
Recombinant Viruses ◽  
Multiple Nucleopolyhedrovirus ◽  
Gfp Fluorescence ◽  
Occlusion Bodies ◽  
Green Fluorescent ◽  
And Control ◽  
Autographa Californica Multiple Nucleopolyhedrovirus

Two envelope fusion protein gene homologues have been identified in the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). AcMNPV GP64 protein is fusogenic and essential for propagation and pathogenicity. The F homologue (Ac23) is not essential, is fusion-incompetent in standard assays, but contributes to faster host death. Here, we show that occlusion bodies (OBs) from Ac23null mutants and control viruses do not differ significantly in size and the number of occlusion-derived virions (ODVs) contained; however, Ac23null OBs had a much higher percentage of ODVs with a single nucleocapsid (44.6 %) than the near-isogenic control (11.3 %). Infection of Sf9 cells with Ac23–green fluorescent protein (gfp)-expressing recombinant viruses showed Ac23–gfp fluorescence overlapping perinuclear DAPI staining at later times, a pattern not observed with GP64. These results suggest that F proteins have evolved functions beyond envelope fusion and play a different role from that of GP64 in viruses that contain both proteins.

CELL PROLIFERATION IN THE SPRUCE BUDWORM, CHORISTONEURA FUMIFERANA (LEPIDOPTERA: TORTRICIDAE), INFECTED WITH ENTOMOPOXVIRUS

1976 ◽  
Vol 108 (8) ◽  
pp. 859-864 ◽  
Author(s):  
F. T. Bird
Keyword(s):  
Cell Proliferation ◽  
Fat Body ◽  
Choristoneura Fumiferana ◽  
Spruce Budworm ◽  
Advanced Stage ◽  
Subsequent Infection ◽  
Large Size

AbstractThe last larval stadium of the spruce budworm, Choristoneura fumiferana (Clemens), is extended, if infection by an entomopoxvirus is in an advanced stage at the normal time of pupation. The larvae attain an abnormally large size. Part of this increase in size is caused by a proliferation and subsequent infection and swelling of fat body cells.

Some cytological changes in the fat body and testes of male spruce budworm (Choristoneura fumiferana Clem.) pupae after treatment with tepa

1971 ◽  
Vol 49 (6) ◽  
pp. 831-836 ◽  
Author(s):  
Norton D. Addy
Keyword(s):  
Protein Metabolism ◽  
Protein Concentration ◽  
Fat Body ◽  
Choristoneura Fumiferana ◽  
Histological Study ◽  
Spruce Budworm ◽  
Protein Patterns ◽  
Dominant Lethal Mutation ◽  
Cytological Changes ◽  
Fat Bodies

A histological study was conducted to determine the effects of tepa, an aziridine-alkylating agent, on male spruce budworm pupae. The results indicated an increase in protein concentration in the fat body and testes of the tepa-treated insects. The profound changes in protein metabolism in the tepa-treated insects are similar to known protein patterns in fat bodies that are mediated by increases in ecdysone titer during the pupal period. These changes are further evident in the eye and testes pigment production and in the differentiating sperm that result in sterile gametes which all depend on ecdysone titer for their normal development. From this it seems evident that tepa may have an effect on the cell's ability to regulate hormone levels, protein metabolism, or both. And these effects result in the critical changes in mature sperm that lead to the dominant lethal mutation occurring in resulting zygotes.

scholarly journals A study of the Autographa californica multiple nucleopolyhedrovirus ODV envelope protein p74 using a GFP tag

2001 ◽  
Vol 82 (9) ◽  
pp. 2279-2287 ◽  
Author(s):  
Jeffrey M. Slack ◽  
Edward M. Dougherty ◽  
Susan D. Lawrence
Keyword(s):  
Spodoptera Frugiperda ◽  
Fluorescent Protein ◽  
Protein Localization ◽  
Chimeric Proteins ◽  
Ring Zone ◽  
Multiple Nucleopolyhedrovirus ◽  
C Terminus ◽  
Green Fluorescent ◽  
Autographa Californica Multiple Nucleopolyhedrovirus ◽  
Gfp Tag

The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) protein p74 is associated with the occlusion-derived virus (ODV) envelope. p74 is essential for oral infectivity of ODV and has been proposed to play a role in midgut attachment and/or fusion. In this study, p74 protein was expressed in-frame with green fluorescent protein (GFP) to create a p74–GFP chimera. The C-terminal GFP portion of the chimera facilitated visualization of the trafficking of p74 in baculovirus-infected Spodoptera frugiperda (Sf-9) cells. p74–GFP chimeric proteins localized in the intranuclear ring zone of the nucleus and were found to co-precipitate with the microvesicle fraction of cell lysates. A series of truncations of p74 was expressed as p74–GFP chimeras in recombinant baculoviruses. When C-terminal region S580–F645 was deleted from p74, p74–GFP chimera localization became non-specific and chimeras became soluble. p74 region S580–F645 directed GFP to the intranuclear ring zone in a similar pattern to full-length p74. The hydrophobic C terminus of p74 plays a role in protein localization and possibly in transmembrane anchoring and insertion.

FEEDING BEHAVIOUR OF THE FIRST-INSTAR CHORISTONEURA FUMIFERANA AND CHORISTONEURA PINUS PINUS (LEPIDOPTERA: TORTRICIDAE)

1999 ◽  
Vol 131 (1) ◽  
pp. 79-84 ◽  
Author(s):  
A. Retnakaran ◽  
W.L. Tomkins ◽  
M.J. Primavera ◽  
S.R. Palli
Keyword(s):  
Fluorescent Protein ◽  
Choristoneura Fumiferana ◽  
Glass Tube ◽  
Choristoneura Pinus ◽  
Jack Pine Budworm ◽  
Occlusion Bodies ◽  
Early Instar ◽  
Choristoneura Pinus Pinus ◽  
Green Fluorescent ◽  
Red Dye

AbstractWe have discovered that, contrary to the long-held belief, 1st-instar spruce budworm, Choristoneura fumiferana Clemens, do feed. They display red alimentary tracts if they are provided with diet containing the red dye amaranth. They graze on the surface of balsam fir needles sprayed with rhodamine and ingest the fluorescent material, which can be detected in the frass pellets deposited inside the hibernacula. When emerging 1st instars were allowed to crawl on the inside surface of a glass tube coated with the polyhedral inclusion bodies of a recombinant C. fumiferana virus containing the gene for the green fluorescent protein, the larvae showed the characteristic green fluorescence, indicating that not only had they ingested the occlusion bodies but also the virus had replicated and infected different tissues. Similar results were obtained with the jack pine budworm, Choristoneura pinus pinus Freeman, which has an identical life history. The advantages of early-instar intervention to minimize defoliation by using control agents such as the ecdysteroid agonist, tebufenozide (RH-5992, Mimic® formulation), are discussed.

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