Abstract
The orf8 gene (Bm8) in Bombyx mori nucleopolyhedrovirus (BmNPV) is one of 17 genes unique to group I NPVs and is expressed as an early gene. We have reported that Bm8 may play an important role during viral infection and that Bm8 protein co-localized with IE1 to specific nuclear foci throughout infection. It was also demonstrated that both IE1 and BmNPV hr facilitate this localization of Bm8. To investigate further, host proteins interacting with Bm8 were screened using a yeast two-hybrid system. We identified 6 host clones as Bm8-interacting partners from three cDNA libraries derived from BmN cells or B. mori larvae. Further assays showed that the N-terminal region of Bm8 is important for the interaction with most host clones and that two of the clones can associate with IE1. Cloning and sequencing of full-length cDNAs revealed that most of the clones potentially encode either membrane-bound proteins or secreted proteins. Quantitative RT-PCR analysis revealed that some of these host genes were slightly induced during the early stage of infection in BmN cells, and that the expression of all genes was markedly reduced during the late stage of infection. Generation of mutant BmNPVs over-expressing these host genes also identified a gene that potentially functions as a negative factor during BmNPV infection. These features of Bm8-interacting host proteins strongly support that Bm8 is a multifunctional protein involved in multiple signaling pathways in host cells.
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References
Beniya, H., Braunagel, S.C., and Summers, M.D. 1998. Autographa californica nuclear polyhedrosis virus: subcellular localization and protein trafficking of BV/ODVE26 to intranuclear membranes and viral envelopes. Virology240, 64–75.
Braunagel, S.C., Cox, V., and Summers, M.D. 2009. Baculovirus data suggest a common but multifaceted pathway for sorting proteins to the inner nuclear membrane. J. Virol.83, 1280–1288.
Braunagel, S.C., Williamson, S.T., Saksena, S., Zhong, Z., Russell, W.K., Russell, D.H., and Summers, M.D. 2004. Trafficking of ODV-E66 is mediated via a sorting motif and other viral proteins: facilitated trafficking to the inner nuclear membrane. Proc. Natl. Acad. Sci. USA101, 8372–8377.
Burks, J.K., Summers, M.D., and Braunagel, S.C. 2007. BV/ODV-E26: a palmitoylated, multifunctional structural protein of Autographa californica nucleopolyhedrovirus. Virology361, 194–203.
Hayward, S.D., Liu, J., and Fujimuro, M. 2006. Notch and Wnt signaling: mimicry and manipulation by gamma herpesviruses. Sci. STKE335, re4.
Herniou, E.A., Luque, T., Chen, X., Vlak, J.M., Winstanley, D., Cory, J.S., and O’Reilly, D.R. 2001. Use of whole genome sequence data to infer baculovirus phylogeny. J. Virol.75, 8117–8126.
Herniou, E.A., Olszewski, J.A., Cory, J.S., and O’Reilly, D.R. 2003. The genome sequence and evolution of baculoviruses. Annu. Rev. Entomol.48, 211–234.
Imai, N., Kurihara, M., Matsumoto, S., and Kang, W.K. 2004. Bombyx mori nucleopolyhedrovirus orf8 encodes a nucleic acid binding protein that colocalizes with IE1 during infection. Arch. Virol.149, 1581–1594.
Jehle, J.A., Blissard, G.W., Bonning, B.C., Cory, J.S., Herniou, E.A., Rohrmann, G.F., Theilmann, D.A., Thiem, S.M., and Vlak, J.M. 2006. On the classification and nomenclature of baculoviruses: a proposal for revision. Arch. Virol.151, 1257–1266.
Kang, W., Imai, N., Kawasaki, Y., Nagamine, T., and Matsumoto, S. 2005. IE1 and hr facilitatethe localization of Bombyx mori nucleopolyhedrovirus ORF8 to specific nuclear sites. J. Gen. Virol.86, 3031–3038.
Kang, W., Imai, N., Suzuki, M., Iwanaga, M., Matsumoto, S., and Zemskov, E.A. 2003. Interaction of Bombyx mori nucleopolyhedrovirus BRO-A and host cell protein LAMININ. Arch. Virol.148, 99–113.
Katsuma, S., Kawaoka, S., Mita, K., and Shimada, T. 2008. Genome-wide survey for baculoviral host homologs using the Bombyx genome sequence. Insect Biochem. Mol. Biol.38, 1080–1086.
Katsuma, S. and Shimada, T. 2009. Bombyx mori nucleopolyhedrovirus ORF34 is required for efficient transcription of late and very late genes. Virology392, 230–237.
Ko, R., Okano, K., and Maeda, S. 2000. Structural and functional analysis of the Xestia c-nigrum granulovirus matrix metalloproteinase. J. Virol.74, 11240–11246.
Maeda, S. 1989a. Expression of foreign genes in insects using baculovirus vectors. Annu. Rev. Entomol.34, 351–372.
Maeda, S. 1989b. Gene transfer vectors of a baculovirus, Bombyx mori, and their use for expression of foreign genes in insect cells. pp. 167–181. In Mitsuhashi, J. (ed.), Invertebrate cell system applications. CRC Press Inc., Boca Raton, FL, USA.
Maeda, S. 1995. Further development of recombinant baculovirus insecticides. Curr. Opin. Biotechnol.6, 313–319.
May, P.E., Woldt, E., Matz, R.L., and Boucher, P. 2007. The LDL receptor-related protein (LRP) family: an old family of proteins with new physiological functions. Ann. Med.39, 219–228.
Mita, K., Kasahara, M.S., Sasaki, S., Nagayasu, Y.T., Yamada, T., Kanamori, H., Namiki, N., Kitagawa, M., Yamashita, H., Yasukochi, Y., andet al. 2004. The genome sequence of silkworm, Bombyx mori. DNA Res.11, 27–35.
Nichols, J.T., Miyamoto, A., and Weinmaster, G. 2007. Notch signaling — constantly on the move. Traffic8, 959–969.
Nie, Y., Fang, M., and Theilmann, D.A. 2009. AcMNPV AC16 (DA26, BV/ODV-E26) regulates the levels of IE0 and IE1 and binds to both proteins via a domain located within the acidic transcriptional activation domain. Virology385, 484–495.
Nie, Y. and Theilmann, D.A. 2010. Deletion of AcMNPV AC16 and AC17 results in delayed viral gene expression in budded virus infected cells but not transfected cells. Virology404, 168–179.
O’Reilly, D.R., Passarelli, A.L., Goldman, I.F., and Miller, L.K. 1990. Characterization of the DA26 gene in a hypervariable region of the Autographa californica nuclear polyhedrosis virus genome. J. Gen. Virol.71, 1029–1038.
Saito, K., Chen, M., Bard, F., Chen, S., Zhou, H., Woodley, D., Polischuk, R., Schekman, R., and Malhotra, V. 2009. TANGO1 facilitates cargo loading at endoplasmic reticulum exit sites. Cell136, 891–902.
Thomas, U., Speicher, S.A., and Knust, E. 1991. The Drosophila gene Serrate encodes an EGF-like transmembrane protein with a complex expression pattern in embryos and wing discs. Development111, 749–761.
Xia, Q., Guo, Y., Zhang, Z., Li, D., Xuan, Z., Li, Z., Dai, F., Li, Y., Cheng, D., Li, R., andet al. 2009. Complete resequencing of 40 genomes reveals domestication events and genes in silkworm (Bombyx). Science326, 433–436.
Xia, Q., Zhou, Z., Lu, C., Cheng, D., Dai, F., Li, B., Zhao, P., Zha, X., Cheng, T., Chai, C., andet al. 2004. A draft sequence for the genome of the domesticated silkworm (Bombyx mori). Science306, 1937–1940.
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Kang, W., Katsuma, S., Matsuda-Imai, N. et al. Identification and characterization of host factors interacting with Bombyx mori nucleopolyhedrovirus ORF8. J Microbiol. 50, 469–477 (2012). https://doi.org/10.1007/s12275-012-2010-z
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DOI: https://doi.org/10.1007/s12275-012-2010-z