Abstract
A baculovirus polyhedrin protein has proven to possess a nuclear localization signal (NLS) sequence and a domain required for supramolecular assembly. Here we investigated five Bombyx mori nucleopolyhedrovirus (BmNPV) mutants that did not produce polyhedra. Two of five mutants were generated during routine baculoviral expression vector screening, and three were isolated by treatment with the mutagen 5-bromo-2′-deoxyuridine (BrdU). Marker rescue mapping and nucleotide sequence analysis showed that mutations in the polyhedrin gene caused the altered phenotype of these mutants. Biochemical fractionation indicated that cells infected with these mutants exhibited polyhedrin protein in both the nucleus and the cytoplasm. Electron microscopic observation revealed that polyhedrin produced by these mutants ocurred in both the nucleus and the cytoplasm, but did not form a crystalline lattice. Despite the incompleteness of polyhedrin nuclear localization, the NLSs of the five mutants were unchanged, although some of the mutations occurred within residues just outside of the domain reported to be required for polyhedron assembly (4). This result suggests that (a) the polyhedrin NLS directs polyhedrin to the nucleus, but the efficiency of this localization is regulated by regions other than the NLS (probably, polyhedrin conformation and its association with the nucleus are also involved), and (b) formation of a crystalline lattice may also be determined by several domains within polyhedrin.
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References
Maeda S., In Insect Cell Biotechnology (K. Maramorosch, and A. McIntosh, eds), CRC Press, Boca Raton, FL., 1994 pp. 1-31.
Choudary P.V., Kamita S.G., and Maeda S., In Methods in Molecular Biology — Baculovirus Expression Protocols (C.D. Richardson, ed), Humana Press, Totowa, NJ, 1995 pp. 243–264.
Miller L.K., J Invertebr Pathol 65, 211–216, 1995.
Jarvis D.L., Bohlmeyer D.A., and Garcia A. Jr., Virology 185, 795–810, 1991.
Eason J.E., Hice R.H., Johnson J.J., and Federici B.A., J Virol 72, 6237–6243, 1998.
Zhou C.E., Ko R., and Maeda S., Virology 240, 282–294, 1998.
Brown N., Faulkner P., and Chung K.L., J Gen Virol 50, 309–316, 1980.
Carstens E.B., Lin-bai Y., and Faulkner P., Virology 68, 901–905, 1987.
Carstens E.B., Williams G.V., Faulkner P., and Partington S., J Gen Virol 73, 1471–1479, 1992.
Noguchi Y., Jpn Appl Entomol Zool 34, 279–282, 1990.
Noguchi Y., J Seric Sci Jpn 60, 302–309, 1991.
Shimada T., Chan I.-C., Noguchi Y., Nagata M., and Kobayashi M., J Seric Sci Jpn 63, 353–360, 1994.
Katsuma S., Noguchi Y., Zhou C.E., Kobayashi M., and Maeda S., J Gen Virol 80, 783–791, 1999.
Katsuma S., Noguchi Y., Shimada T., Nagata M., Kobayashi M., and Maeda S., Arch Virol 144, 1275–1285, 1999.
Maeda S., Kawai T., Obinata M., Fujiwara H., Horiuchi T., Saeki Y., Sato Y., and Furusawa M., Nature, London 315, 592–594, 1985.
Maeda S. and Majima K., J Gen Virol 71, 1851–1855, 1990.
Majima K., Kobara R., and Maeda S., J Virol 67, 7513–7521, 1993.
Jarvis D.L., Bohlmeyer D.A., and Garcia A. Jr., J Virol 66, 6903–6911, 1992.
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Katsuma, S., Deng, D.Xt., Ecale Zhou, C.L. et al. Identification of Novel Residues Involved in Nuclear Localization of a Baculovirus Polyhedrin Protein. Virus Genes 21, 233–240 (2000). https://doi.org/10.1023/A:1008151916849
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DOI: https://doi.org/10.1023/A:1008151916849