Summary
Using immuno-electron microscopy the 34 kDa polyhedron envelope (PE) phosphoprotein (pp 34) was localized in cells infected withAutographa californica multiple-nucleocapsid nuclear polyhedrosis virus (AcMNPV). In wild-type AcMNPV-infected cells this protein was found associated with electron-dense “spacers” and the polyhedron envelopes demonstrating their structural relationship. In these cells pp 34 was also found associated with fibrillar structures present in the nucleus and cytoplasm of infected cells. However, when cells were infected with an AcMNPV mutant with an inactivated pp 34 gene, antiserum against pp 34 still localized in fibrillar structures. In cells infected with AcMNPV mutants lacking p 10, and thus devoid of fibrillar structures, pp 34 localized normally with both electron-dense “spacers” and polyhedron envelopes. These data confirm that fibrillar structures are not essential for the morphogenesis of polyhedron envelopes and demonstrate that the association of anti pp 34 serum with these structures is specific but fortuitous.
Similar content being viewed by others
References
Bicknell JN, Leisy DJ, Rohrmann GF, Beaudreau GS (1987) Comparison of the p 26 gene region of two baculoviruses. Virology 161: 589–592
Croizier G, Gonnet P, Devauchelle G (1987) Localization cytologique de la protéine non structurale p 10 du baculovirus de la polyédrose nucléaire de LépidoptèreGalleria mellonella L. C Rend Acad Sci 305: 677–681
Faulkner P (1981) Baculoviruses. In: Davidson EA (ed) Pathogenesis of invertebrate microbial diseases. Allanheld, Osmun & Co, Totowa NJ, pp 3–37
Gombart AF, Blissard GW, Rohrmann GF (1989a) Characterization of the genetic organization of the HindIII-M region of the multicapsid nuclear polyhedrosis virus ofOrgyia pseudostugata reveals major differences among baculoviruses. J Gen Virol 70: 1815–1828
Gombart AF, Pearson MN, Rohrmann GF, Beaudreau GS (1989b) A baculovirus PE-associated protein: genetic location, nucleotide sequence and immunocytochemical characterization. Virology 169: 182–193
Gonnet P, Devauchelle G (1987) Obtention par recombinaison dans le gène du polypeptide p 10 d'un baculovirus exprimant le gène de résistance à la néomycine dans les cellules d'insecte. C Rend Acad Sci 305: 111–114
Granados RR, Federici BA (eds) (1986) The biology of baculoviruses, 2 vols. CRC Press, Boca Raton, FL
Hink F (1970) Established insect cell line from the cabbage looper,Trichoplusia ni. Nature 226: 466–467
Kelly DC (1982) Baculovirus replication. J Gen Virol 63: 1–13
Kormelink RM, Jore JJ, Usmany M, Klinge-Roode EC, van Lent JWM, Vlak JM (1989) Engineering ofAutographa californica nuclear polyhedrosis virus expression vectors using the p 10 promoter. In preparation
Kushida H (1961) A new embedding method for ultrathin sectioning using a methacrylate resin with three dimensional polymer structure. J Microsc 10: 194–197
Kuzio J, Rohel DZ, Curry CJ, Krebs A, Carstens EH, Faulkner P (1984) Nucleotide sequence of the p 10 polypeptide gene ofAutographa californica nuclear polyhedrosis virus. Virology 139: 414–418
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Leisy DJ, Rohrmann GF, Beaudreau GS (1984) Conservation of genome organization in two multicapsid nuclear polyhedrosis viruses. J Virol 52: 699–702
Luckow VL, Summers MD (1988) Trends in the development of baculovirus expression vectors. Bio/Technol 6: 47–55
MacKinnon EA, Henderson JF, Stoltz DB, Faulkner P (1973) Morphogenesis of nuclear polyhedrosis virus under conditions of prolonged passage in vitro. J Ultrastruct Res 49: 419–435
Maeda S (1989) Expression of foreign genes in insects using baculovirus vectors. Ann Rev Entomol 34: 351–372
Martignoni ME (1984) Baculovirus: an attractive alternative. In: Garner WY, Harvey J (eds) Chemicals and biological controls in agriculture and forestry. American Chemical Society, Washinton, DC, pp 51–67
Miller LK (1988) Baculoviruses as gene expression vectors. Ann Rev Microbiol 42: 177–199
Minion FC, Coons LB, Broome JR (1979) Characterization of the PE of the nuclear polyhedrosis virus ofHeliothis virescens. J Invertebr Pathol 34: 303–307
Oellig C, Happ B, Müller T, Doerfler W (1987) Overlapping sets of viral RNAs reflect the array of polypeptides in theEcoRI J and N fragments (map positions 81.2 to 85.0) of theAutographa california nuclear polyhedrosis virus. J Virol 61: 3048–3057
Oellig C, Happ B, Müller T, Doerfler W (1989) Overlapping sets of viral RNAs reflect the array of polypeptides in theEcoRI J and N fragments (map positions 81.2 to 85.0) of theAutographa californica nuclear polyhedrosis virus. J Virol 63: 1494
Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17: 208–212
Russell RLQ, Rohrmann GF (1990) Baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra. Virology 174 (in press)
Smith GE, Summers MD (1978) Analysis of baculovirus genomes with restriction endonucleases. Virology 98: 517–527
Smith GE, Summers MD (1981) Application of a novel radioimmunoassay to identify baculovirus structural proteins that share interspecies antigenic determinants. J Virol 39: 125–137
Smith GE, Vlak JM, Summers MD (1983) Physical analysis ofAutographa californica nuclear polyhedrosis virus transcripts for polyhedrin and 10,000-molecular weight protein. J Virol 45: 215–225
van der Wilk F, van Lent JWM, Vlak JM (1987) Immunogold detection of polyhedrin, p 10 and virion antigens inAutographa californica nuclear polyhedrosis virus-infectedS. frugiperda cells. J Gen Virol 68: 2615–2623
van Lent JWM, Verduin BJM (1986) Detection of viral protein and particles in thin sections of infected plant tissue using immunogold labeling. In: Jones RAC, Torrance L (eds) Developments in applied biology, vol 1, developments and applications in virus testing. Assoc Applied Biologists, Wellesborne/Levenham Press, London, pp 193–211
Vaughn JL, Goodwin RH, Tompkins GJ, McCawley P (1977) The establishment of two cell lines from the insectSpodoptera frugiperda (Lepidoptera: Noctuidae). In Vitro 13: 213–217
Vlak JM (1979) The proteins of nonoccludedAutographa californica nuclear polyhedrosis virus produced in an established cell line ofSpodoptera frugiperda. J Invertebr Pathol 34: 110–118
Vlak JM, Klinkenberg FA, Zaal KJM, Usmany M, Klinge-Roode EC, Geervliet JBF, Roosien J, van Lent JWM (1988) Functional studies on the p 10 gene ofAutographa californica nuclear polyhedrosis virus using a recombinant expressing a p 10-β-galactosidase fusion gene. J Gen Virol 69: 765–776
Whitt MA, Manning JS (1988) A phosphorylated 34-kDa protein and a subpopulation of polyhedrin are thiol linked to the carbohydrate layer surrounding a baculovirus occlusion body. Virology 163: 33–42
Williams GV, Rohel DZ, Kuzio J, Faulkner P (1989) A cytopathological investigation ofAutographa californica nuclear polyhedrosis virus p 10 gene function using insertion/deletion mutants. J Gen Virol 70: 187–202
Zuidema D, Klinge-Roode EC, Van Lent JWM, Vlak JM (1989) Construction and analysis of anAutographa californica nuclear polyhedrosis virus mutant lacking the polyhedral envelope. Virology 173: 98–108
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
van Lent, J.W.M., Groenen, J.T.M., Klinge-Roode, E.C. et al. Localization of the 34 kDa polyhedron envelope protein inSpodoptera frugiperda cells infected withAutographa california nuclear polyhedrosis virus. Archives of Virology 111, 103–114 (1990). https://doi.org/10.1007/BF01310508
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01310508