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Insect |
Department of Plant and Microbial Biology, University of California, 251 Koshland Hall, Berkeley, CA 94720-3102, USA1
Author for correspondence: Loy Volkman. Fax +1 510 642 4995. e-mail lvolkman{at}nature.berkeley.edu
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Abstract |
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Introduction |
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TOPAbstractIntroduction Methods Results Discussion References |
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). This diversity in hosts is echoed by diversity among the baculoviruses themselves, and is reflected not only by differences in the host range of individual viruses, but also in pathogenesis, variation in genome size and G+C content (Miller, 1997 ; Ahrens et al., 1997 ). Baculovirus genomes are large and contain over 100 open reading frames, but only a small fraction of these have been functionally characterized even for the best-studied baculovirus, Autographa californica M nucleopolyhedrovirus (AcMNPV) (Miller, 1997 ). AcMNPV, the type-species of the Nucleopolyhedrovirus genus, has served as the de facto model for all baculoviruses even though it may not be representative of the group as a whole, or even, for that matter, of the Nucleopolyhedrovirus genus. For example, AcMNPV produces two different forms to complete its infection cycle in larval lepidopteran hosts. One form, released from an occluding protein matrix following ingestion by a susceptible host, initiates infection in midgut cells while the other form, budded virus (BV), transmits infection systemically. The major envelope glycoprotein of AcMNPV BV, gp64, is encoded by the AcMNPV genome and is essential for functional entry of AcMNPV BV. The recent completion of the genomic sequences of two other nucleopolyhedroviruses, Lymantria dispar (Ld)MNPV (Kuzio et al., 1999 ) and Helicoverpa zea (Hz)SNPV (Albert Lu, DuPont Agricultural Products, personal communication), however, has revealed that homologues of this essential AcMNPV gene are missing in those genomes. This finding leaves a void in our understanding of how the budded forms of these two viruses functionally enter their host cells, and also serves to remind us that what may be true for AcMNPV may not be true for every baculovirus.
Given the uncertainties of AcMNPV as a representative of other nucleopolyhedroviruses, we were anxious to determine whether our recent finding, that filamentous actin (F-actin) is essential for nucleocapsid morphogenesis of AcMNPV (Ohkawa & Volkman, 1999 ), holds true for other nucleopolyhedroviruses. The involvement of F-actin is quite unexpected and unique for viruses that replicate and assemble within the nucleus; hence, an understanding of the breadth of the requirement among baculoviruses is fundamental to any understanding of baculovirus–host interactions as a whole. Further, the implications are that baculoviruses that are found to be F-actin dependent must encode gene products that specifically interact with, and manipulate, the actin cytoskeleton of their hosts in an (as yet) unprecedented way.
Our study was conducted by testing the ability of six divergent members of the Nucleopolyhedrovirus genus to produce progeny in the presence of two actin-binding drugs, cytochalasin D (CD) and latrunculin A (LA), which differ in their modes of action. The viruses we used were selected because cloned isolates and cell lines for their propagation were available, and because phylogenetic studies based on two conserved, virus-encoded proteins (polyhedrin and DNA polymerase) indicated that this set of lepidopteran viruses spanned the Nucleopolyhedrovirus genus with regard to evolutionary relatedness (Zanotto et al., 1993 ; Hu, 1998 ; Bulach et al., 1999 ). Notably, we included the two nucleopolyhedroviruses known not to encode gp64. Here we present results demonstrating that for all six, progeny production was sensitive to both CD and LA, suggesting that F-actin is a genus-wide requirement for the replication of those nucleopolyhedroviruses with lepidopteran hosts.
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Methods |
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TOPAbstractIntroduction Methods Results Discussion References |
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Cell lines and media. ). Bombyx mori BmN cells (provided by S. George Kamita at the University of California, Davis, CA, USA) were grown in TC-100 medium and maintained as described (Maeda, 1989 ). Heliothis virescens-derived BCIRL-HV-AM1 cells (McIntosh et al., 1981 ) were provided by James Wong (DuPont Agricultural Products, Newark, DE, USA) and grown in TNMFH medium with 10% foetal bovine serum. Sf-9, Sf-21, TN-368 and BCIRL-HV-AM1 cells were sometimes grown in suspension cultures, at which time their media were supplemented with Pluronic acid F-68 (Sigma) to 0·1%.
Viruses.
All viruses used were cloned isolates and included AcMNPV-hsp70/lacZ (Engelhard et al., 1994 ), Spodoptera frugiperda (Sf)MNPV, Bombyx mori (Bm)NPV, Orgyia pseudotsugata (Op)MNPV, LdMNPV, Anticarsia gemmatalis (Ag)MNPV and HzSNPV-hsp70/lacZ. AcMNPV-hsp70/lacZ was a first-passage stock amplified in Sf-9 cells after inoculation with infectious H. virescens haemolymph. BmNPV budded virus stock was the gift of S. George Kamita. OpMNPV budded virus was generously provided by George Rohrmann. Susan Thiem (Michigan State University, East Lansing, MI, USA) and James Slavicek (USDA Forest Service, Delaware, OH, USA) both provided budded virus stocks of LdMNPV A21 (Slavicek et al., 1996 ). AgMNPV-ag3 and SfMNPV-clone 1 were obtained from D. E. Lynn. HzSNPV-hsp70/lacZ was provided by James Wong.
Drug treatments.
CD (Sigma) and LA (Molecular Probes) were dissolved in dimethylsulfoxide at a minimum concentration of 500x and frozen in aliquots at -80 °C until needed. Cells from exponential phase cultures (8x105 cells/ml for cells grown in flasks and 1·2x106 cells/ml for suspension cultures) were seeded at 106 per well in six-well plates (Falcon) and allowed to settle for 1–2 h. The exception was HvE1a cells, which were passaged by trypsinization. These cells were seeded at one-half density and allowed to settle for 24 h in order to recover. The media were aspirated from the wells, and equal amounts of inocula were added at an m.o.i. of 10–20. The m.o.i.s used with LdMNPV and SfMNPV were lower because of lower titres (2 and 0·1, respectively). The six-well plates were rocked slowly at room temperature for a 1 h adsorption period, after which the inocula were removed and the cells washed gently twice with fresh medium. Each well then received 2 ml of fresh medium, with or without CD or LA. This medium was immediately sampled (30 µl, or 100 µl for SfMNPV and LdMNPV) for baseline (residual) BV titres. Additional samples were taken at 24, 48 and sometimes 72 h post-infection (p.i.). All samples were frozen at -80 °C immediately after they were taken, and kept frozen until they were titred by immunoplaque assay. Each experiment was done independently twice.
Plaque assays.
Immunoplaque assays were performed essentially as previously described (Volkman & Goldsmith, 1982 ), with modifications specific to each virus (Table 1). Briefly, all assays were done with microscope slides that were embossed with twelve 5 mm circular wells per slide (Carlson Scientific no. 101205). The slides were prepared for cell culture by immersing them overnight in glass staining-dishes containing a solution of 70% ethanol and 1% HCl, followed by 10 rinses in tap-water and then glass-distilled water. Dishes containing rinsed slides were covered and baked overnight at 75 °C and used without further sterilization. Prepared slides were placed in sterile, covered, plastic dishes (Falcon no. 1012) and seeded with a subconfluent monolayer of actively growing cells. After the cells had settled for 30 min to 1 h, the medium was removed and replaced with 10 µl of a serial dilution of sampled cell supernatant, and the slide incubated at 27 °C. After 1 h, the inoculum was aspirated and quickly replaced with 0·6% hydroxypropylmethylcellulose (Dow Chemical) dissolved in the appropriate medium. Incubation at 27 °C was then continued for 40–96 h, depending on the virus.
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-D-galactopyranoside in 5 mM K4Fe(CN)6, 5 mM K3Fe(CN)6, 2 mM MgCl2 in distilled water] and incubated for 1–2 h to develop the signal. For the remaining viruses, cells were blocked with dilute normal goat serum for 5 min after fixing and rinsing, and then incubated with the appropriate primary antibody for 25 min (Table 1
). Antibodies used for this purpose were rabbit antibodies to AcMNPV polyhedrin (Volkman, 1983 ), OpMNPV MBP–ORF2 (Russell et al., 1997 ; generously provided by G. F. Rohrmann) and mouse monoclonal antibody to AcMNPV gp64 (Keddie et al., 1989 ). After several PBS rinses, a secondary horseradish peroxidase-conjugated antibody was incubated with the cells for 25 min. After a final rinse, True Blue Peroxidase substrate (Kirkegaard and Perry Laboratories) was overlaid to produce a blue stain at the locations of plaques. Each slide was then air-dried, mounted in D. P. X. (Aldrich), and the plaques counted using a compound microscope equipped with a 40x objective. Each serial dilution was plated in quadruplicate, and each relevant dilution was assayed twice.
Western blots.
Suspension cultures of Sf-9, Sf-21 and TN-368 cells in exponential phase growth, all growing in TNMFH+10% FBS+0·1% Pluronic acid, were seeded into duplicate 35 mm plates at 106 cells per plate. Cells were allowed to settle and spread for 12 h, and then they were rinsed twice quickly with PBS, drained, and lysed in 150 µl cell lysis buffer (PBS+0·5% NP-40). Plates were scraped with a cell scraper, and the lysate was placed in microfuge tubes (one per plate) on ice. Fifty µl of 4x SDS–PAGE sample buffer containing E64 proteinase inhibitor (final concentration 30 µg/ml) was added to each tube (Hom & Volkman, 1998 ), and the contents were then immediately boiled for 2 min, chilled on ice briefly, and stored at -80 °C. Thawed samples were diluted 1:2 in series up to 1:64 in sample buffer (1x), boiled again for 1 min, and then loaded onto 10% acrylamide SDS–PAGE gels. After electrophoresis, the proteins were transferred to Immobilon-P membrane (Millipore) and probed with C4 anti-actin antibody (Lessard, 1988 ) diluted 1:2000 in TBS containing 0·2% Tween 20, followed by goat anti-mouse horseradish peroxidase-conjugated antibody diluted 1:3000 in the same diluent (Sigma). Reactions were elucidated using the Supersignal Chemiluminescent Substrate Luminol/Enhancer (Pierce).
Immunofluorescence microscopy.
Approximately 105 actively growing TN-368 cells in 200 µl medium were seeded onto 22 mm square coverslips which had been prepared in the same way as the microscope slides described under Plaque assays. Cells were allowed to settle for 30 min and were then inoculated (where indicated) with AcMNPV-hsp70/lacZ at an m.o.i. of 50. After 1 h, the inocula were removed and replaced with 1 ml fresh medium containing either no drug, 1 µg/ml (2 mM) CD or 0·4 µg/ml (1 mM) LA. After incubation for 25 h at 27 °C, these cells were fixed and treated with monoclonal antibody 39P10 (to AcMNPV p39) for confirmation of infection, TRITC–phalloidin for F-actin staining and DAPI for DNA staining to locate the nucleus as described (Charlton & Volkman, 1991 ). The cells were viewed using a Zeiss Axiophot photomicroscope equipped for fluorescence microscopy.
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Results |
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TOPAbstractIntroduction Methods Results Discussion References |
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; Spector et al., 1989 ); CD binds F-actin at the barbed ends and prevents association and dissociation of G-actin monomers from that end (Brown & Spudich, 1981 ). Both drugs interfere with F-actin-dependent processes, however. The effects of these drugs on F-actin in AcMNPV-infected and uninfected TN-368 cells are shown in Fig. 1. In the absence of drug treatment, the redistribution of F-actin from micospikes and the cortical regions of the cells to the nuclei was clearly evident 25 h p.i. (compare Fig. 1A to 1D). Addition of CD (1 µg/ml) to uninfected cells also resulted in a change in the distribution of F-actin from surface structures (Fig. 1A) to aggregates or foci primarily within the cytoplasm at 25 h p.i. (Fig. 1B). In comparison, addition of CD to infected cells did not appear to affect the distribution so much as the content: CD diminished the amount of F-actin apparent within the nucleus, but some small cytoplasmic aggregates were also evident (compare Fig. 1D to 1E). Incubation of cells for 25 h with LA (0·4 µg/ml), whether uninfected or infected, resulted in the disappearance of F-actin.
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), we initiated our study by testing the sensitivity of AcMNPV-hsp70/lacZ BV production to CD and LA in three permissive cell lines, TN-368, Sf-21 and Sf-9 (Fig. 2A). BV production was sensitive to both drugs individually in all three cell lines, but the amount of drug required for a similar level of inhibition differed for each. Among the three cell lines tested, the greatest sensitivity of BV production to CD was observed in TN-368 cells and the least sensitivity in Sf-21 cells, even though Western blot analysis indicated that the actin content of TN-368 and Sf-21 cells, uninfected, was equivalent (Fig. 2B). These differences in sensitivity to CD suggested that AcMNPV-infected Sf-21 cells contained more F-actin barbed ends than AcMNPV-infected TN-368 cells. This could occur, even if the total F-actin content of both cell types was identical, if the filaments were shorter but more numerous in infected Sf-21 cells. Whatever the cause, the results indicate (not unexpectedly) that cellular factors play a role in the number and character of the actin filaments in AcMNPV-infected cells.
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). These results were consistent with, and inversely correlated with, total actin content per cell for the three cell types tested (Fig. 2B). Because the activity of LA is to sequester monomeric actin, the degree of sensitivity of AcMNPV to LA should be an indication of the size of the pools of G-actin available to the virus. The results, therefore, indicated that the size of the pools of G-actin was roughly equivalent within AcMNPV-infected TN-368 and Sf-21 cells, but smaller in Sf-9 cells.
CD and LA inhibit BV production by six divergent nucleopolyhedroviruses
Table 1 lists the viruses used in this study, their phylogenetic groups, the host cell lines used to support their replication and the antibody or reagent used to determine their titre. The sensitivities of six of these viruses to CD and LA are shown in Figs 3 and 4, respectively. BV production for all six was sensitive to both drugs individually suggesting that they all were dependent upon F-actin for progeny virus production. Infections by all six baculoviruses were completely inhibited by the lowest concentration of CD used, 0·5 µg/ml (Fig. 3). Replication of one of the six, SfMNPV, took place in Sf-21 cells, the same cells wherein this concentration of CD had very little effect on AcMNPV replication (Fig. 2). This difference in CD sensitivity indicated that AcMNPV-infected Sf-21 cells contained more F-actin barbed ends than SfMNPV-infected Sf-21 cells, which, in turn, suggested that virus-encoded factors also had an effect on the number and character of actin filaments within infected cells.
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with Fig. 4). Similarly, by 72 h p.i., L652Y cells had generated a 112-fold higher level of BV when infected by OpMNPV than by LdMNPV (Fig. 4). Further, AcMNPV progeny production in Sf-21 cells was not significantly affected by the addition 0·08 µg/ml of LA at either 24 or 48 h p.i. while SfMNPV progeny production, by comparison, was reduced by 80% at 24 h p.i. and by 70% at 48 h p.i. (compare Fig. 2 with Fig. 4). Likewise, in the presence of 0·08 µg/ml of LA, OpMNPV progeny production was depressed 16-fold by 72 h p.i. compared with 157-fold for LdMNPV at that time (Fig. 4). Together, these results demonstrated that for each pair of viruses infecting a given cell line, the one that replicated the best was the least sensitive to LA. The implications were that the G-actin pool size was affected by virus replication, and the degree of virus enhancement of the pool size may have been a factor in the relative productivity of infection in these cell lines. Because these NPVs do not encode actin (Ahrens et al., 1997 ) and there is no increase in actin synthesis upon infection (Talhouk & Volkman, 1991 ), the source of G-actin is likely to be depolymerized cytoplasmic F-actin. |
Discussion |
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TOPAbstractIntroduction Methods Results Discussion References |
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), but the functions in question all take place within the cytoplasm with the exception of AcMNPV nucleocapsid morphogenesis (Ohkawa & Volkman, 1999 ). The very existence of nuclear actin has been controversial until recently, but although widely accepted now, unequivocal functions for nuclear actin have been difficult to demonstrate (Gonsior et al., 1999 ). In this study we have shown that like AcMNPV, the type species, progeny BV production by six divergent nucleopolyhedroviruses is inhibited by two drugs that interfere with F-actin function. While specificity is always a concern when drawing conclusions from studies involving drugs, the fact that the activities of the two drugs differ (LA sequesters monomeric actin while CD binds the barbed end of F-actin) supports the conclusion that actin is the functional target of the drugs. Further evidence for this is the inverse correlation of the cellular actin content and the concentration of LA needed to interfere with AcMNPV-hsp70/lacZ BV production (Fig. 2). The fact that a similar correlation could not be drawn for CD is expected as the number of barbed ends of F-actin filaments is dependent upon more factors than simply actin concentration. Factors such as filament length and the number, nature and state of cellular actin-binding proteins could all have an effect on the number of barbed ends available within a cell. Indeed, Lanier & Volkman (1998) determined that AcMNPV capsids and nucleocapsids themselves nucleate the polymerization of F-actin in vitro. They further found that addition of CD to the polymerization reaction has a greater impact in reducing the amount of polymerized actin when the virus components are present than when they are absent, indicating that virus-induced nucleation of actin polymerization occurs via the pointed end of F-actin. These results suggest that within infected cells, the presence of capsids and nucleocapsids could be among the factors positively influencing the number of barbed ends available. They also could explain the diminished staining of F-actin observed in infected TN-368 cells treated with CD (Fig. 1E) compared with control infected cells (Fig. 1D).
The viruses chosen for this study have all been subjected to phylogenetic analysis using amino acid sequence data of conserved proteins inferred from the nucleotide sequences of the genes that encode them (Zanotto et al., 1993 ; Hu, 1998 ; Bulach et al., 1999 ). While the computer programs used to analyse the data differ, and the resulting phylogenetic trees are not identical, there is agreement that there are two major clades or groups of NPVs, I and II. AcMNPV, BmNPV, OpMNPV and AgMNPV all fall within group I, while LdMNPV, HzSNPV and SfMNPV fall within group II (Bulach et al., 1999 ). So far, only MNPVs (viruses having one to many nucleocapsids per virion) have been placed in group I, while both SNPVs (viruses with only a single nucleocapsid per virion) and MNPVs fall in group II. Interestingly, those NPVs lacking gp64 fall in group II. Our evidence suggests that the lepidopteran nucleopolyhedrovirus dependence on F-actin evolved prior to the establishment of the two groups as well as before the development of nucleocapsid packaging differences.
All evidence to date indicates that F-actin plays a role in nucleocapsid assembly of nucleopolyhedroviruses within the nucleus (Volkman et al., 1987 ; Volkman, 1988 ; Hess et al., 1989 ). The ability to commandeer and direct the transport of actin to the nucleus is a feature not shared by any other group of viruses. Whether this sort of manipulation of the actin cytoskeleton is more related to the unique capabilities of nucleopolyhedroviruses or to an unexplored vulnerability of lepidopteran cells remains an open question. It will be interesting in this regard to determine whether the granuloviruses which are restricted to lepidopteran hosts, or those nucleopolyhedroviruses that have non-lepidopteran hosts, also depend on F-actin for progeny production.
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Acknowledgments |
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Footnotes |
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References |
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TOPAbstractIntroduction Methods Results Discussion References |
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Brown, S. S. & Spudich, J. A. (1981). Mechanism of action of cytochalasin: evidence that it binds to actin filament ends.Journal of Cell Biology88, 487-491.
Bulach, D. M., Kumar, C. A., Zaia, A., Liang, B. & Tribe, D. E. (1999). Group II nucleopolyhedrovirus subgroups revealed by phylogenetic analysis of polyhedrin and DNA polymerase gene sequences.Journal of Invertebrate Pathology73, 59-73.[Medline]
Charlton, C. A. & Volkman, L. E. (1991). Sequential rearrangement and nuclear polymerization of actin in baculovirus-infected Spodoptera frugiperda cells.Journal of Virology65, 1219-1227.
Coué, M., Brenner, S. L., Spector, I. & Korn, E. D. (1987). Inhibition of actin polymerization by latrunculin A.FEBS Letters213, 316-318.[Medline]
Cudmore, S., Reckmann, I. & Way, M. (1997). Viral manipulations of the actin cytoskeleton.Trends in Microbiology5, 142-148.[Medline]
Engelhard, E. K., Kam-Morgan, L. N. W., Washburn, J. O. & Volkman, L. E. (1994). The insect tracheal system: a conduit for the systemic spread of Autographa californica M nuclear polyhedrosis virus.Proceedings of the National Academy of Sciences, USA91, 3224-3227.
Gonsior, S. M., Platz, S., Buchmeier, S., Scheer, U. & Jockusch, B. M. (1999). Conformational difference between nuclear and cytoplasmic actin as detected by monoclonal antibody.Journal of Cell Science112, 797-809.[Abstract]
Hess, R. T., Goldsmith, P. A. & Volkman, L. E. (1989). Effect of cytochalasin D on cell morphology and AcMNPV replication in a Spodoptera frugiperda cell line.Journal of Invertebrate Pathology53, 169-182.[Medline]
Hom, L. G. & Volkman, L. E. (1998). Preventing proteolytic artifacts in the baculovirus expression system. BioTechniques24, 18-19.
Hu, Z. (1998). Characterization of the Buzura suppressaria single-nucleocapsid nucleopolyhedrovirus genome: a (phylo)genetic study. Dissertation, Wageningen University, Wageningen, The Netherlands.
Keddie, B. A., Aponte, G. W. & Volkman, L. E. (1989). The pathway of infection of Autographa californica nuclear polyhedrosis virus in an insect host.Science243, 1728-1730.
Kuzio, J., Pearson, M. N., Harwood, S. H., Funk, C. J., Evans, J. T., Slavicek, J. M. & Rohrman, G. F. (1999). Sequence and analysis of the genome of a baculovirus pathogenic for Lymantria dispar.Virology253, 17-34.[Medline]
Lanier, L. M. & Volkman, L. E. (1998). Actin binding and nucleation by Autographa californica M nucleopolyhedrovirus. Virology243, 167-177.[Medline]
Lessard, J. L. (1988). Two monoclonal antibodies to actin: one muscle selective and one generally reactive. Cell Motility and Cytoskeleton10, 349-362.[Medline]
Lynn, D. E. & Shapiro, M. (1998). New cell lines from Heliothis virescens: characterization and susceptibility to baculoviruses. Journal of Invertebrate Pathology72, 276-280.[Medline]
McIntosh, A. H., Andrews, P. A. & Ignoffo, C. M. (1981). Establishment of two continuous cell lines of Heliothis virescens (F.) (Lepidoptera: Noctuidae).In Vitro17, 649-650.
Maeda, S. (1989). Gene transfer vectors of a baculovirus, Bombyx mori nuclear polyhedrosis virus, and their use for expression of foreign genes in insect cells. In Invertebrate Cell System Applications, pp. 167-181. Edited by J. Mitsuhashi. Boca Raton, FL: CRC Press.
Miller, L. K. (1997). The Baculoviruses. New York: Plenum.
Ohkawa, T. & Volkman, L. E. (1999). Nuclear F-actin is required for AcMNPV nucleocapsid morphogenesis.Virology264, 1-4.[Medline]
Russell, R. L. Q., Funk, C. J. & Rohrmann, G. F. (1997). Association of a baculovirus-encoded protein with the capsid basal region. Virology227, 142-152.[Medline]
Slavicek, J. S., Mercer, M. J., Kelly, M. E. & Hayes-Plazolles, N. (1996). Isolation of a baculovirus variant that exhibits enhanced polyhedra production stability during serial passage in cell culture.Journal of Invertebrate Pathology67, 153-160.
Spector, I., Shochet, N. R., Blasberger, D. & Kashman, Y. (1989). Latrunculins–novel marine macrolides that disrupt microfilament organization and affect cell growth. I. Comparison with cytochlasin D.Cell Motility and the Cytoskeleton13, 127-144.[Medline]
Talhouk, S. N. & Volkman, L. E. (1991). Autographa californica M nuclear polyhedrosis virus and cytochalasin D: antagonists in the regulation of protein synthesis.Virology182, 626-634.[Medline]
Volkman, L. E. (1983). Occluded and budded Autographa californica nuclear polyhedrosis virus: immunological relatedness of structural proteins.Journal of Virology46, 221-229.
Volkman, L. E. (1988). Autographa californica MNPV nucleocapsid assembly: inhibition by cytochalasin D.Virology163, 547-553.[Medline]
Volkman, L. E. & Goldsmith, P. A. (1982). Generalized immunoassay for Autographa californica nuclear polyhedrosis virus infectivity in vitro.Applied and Environmental Microbiology 44, 227-233.
Volkman, L. E., Goldsmith, P. A. & Hess, R. T. (1987). Evidence for microfilament involvement in budded Autographa californica nuclear polyhedrovirus virus production.Virology156, 32-39.
Zanotto, P. M. A., Kessing, B. D. & Maruniak, J. E. (1993). Phylogenetic interrelationships among baculoviruses: evolutionary rates and host associations.Journal of Invertebrate Pathology62, 147-164.[Medline]
Received 12 January 2000;
accepted 31 March 2000.
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