The potexvirus Potato virus X(PVX) can be modified genetically to generate chimeric virus particles (CVPS) carrying heterologous peptides fused to coat protein (CP) subunits. A spontaneous PVX mutant expressing a truncated, but functional, form of the CP has been isolated. With the aim of exploiting this virus to display peptides useful for vaccine formulations, two novel viral expression vectors based on pPVX201 (bearing the wild-type PVX genome) were constructed encoding the truncated CP. Both vectors were able to produce infectious virus particles in planta and were used to insert a panel of sequences encoding pepticles of biopharmaceutical interest as N-terminal fusions to the truncated cp gene. The analysis of infection progression induced by the different constructs enabled identification of two important structural features of the fused peptide, namely tryptophan content and isoelectric point, critically affecting the formation of PVX CVPs and virus movement through the plant. These results are discussed in view of the rising interest in engineered plant viruses for development of peptide-based epitope vaccines. © 2006 SGM.

Cited By :35Export Date: 17 July 2015CODEN: JGVIACorrespondence Address: Baschieri, S.; Sezione di Genetica e Genomica Vegetale, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, Italy; email: selene.baschieri@casaccia.enea.itChemicals/CAS: tryptophan, 6912-86-3, 73-22-3; Capsid Proteins; Recombinant Fusion ProteinsReferences: Atabekov, J.G., Rodionova, N.P., Karpova, O.V., Kozlovsky, S.V., Novikov, V.K., Arkhipenko, V., Translational activation of encapsidated potato virus X RNA by coat protein phosphorylation (2001) Virology, 286, pp. 466-474;Baratova, L.A., Grebenshchikov, N.I., Dobrov, E.N., The organization of potato virus X coat proteins in virus particles studied by tritium planigraphy and model building (1992) Virology, 188, pp. 175-180. , & 7 other authors; Baratova, L.A., Grebenshchikov, N.I., Shishkov, A.V., Kashirin, I.A., Radavsky, Y.L., Järvekülg, L., Saarma, M., The topography of the surface of potato virus X: Tritium planigraphy and immunological analysis (1992) J Gen Virol, 73, pp. 229-235; Baratova, L.A., Fedorova, N.V., Dobrov, E.N., N-terminal segment of potato virus X coat protein subunits is glycosylated and mediates formation of a bound water shell on the virion surface (2004) Eur J Biochem, 271, pp. 3136-3145. , & 7 other authors; Baulcombe, D.C., Chapman, S., Santa Cruz, S., Jellyfish green fluorescent protein as a reporter for virus infections (1995) Plant J, 7, pp. 1045-1053; Bendahmane, M., Koo, M., Karrer, E., Beachy, R.N., Display of epitopes on the surface of tobacco mosaic virus: Impact of charge and isoelectric point of the epitope on virus-host interactions (1999) J Mol Biol, 290, pp. 9-20; Borisova, G., Borschukova, O., Skrastina, D., Dislers, A., Ose, V., Pumpens, P., Grens, E., Behavior of a short preS1 epitope on the surface of hepatitis B core particles (1999) Biol Chem, 380, pp. 315-324; Brennan, F.R., Jones, T.D., Longstaff, M., Chapman, S., Bellaby, T., Smith, H., Xu, F., Flock, J.I., Immunogenicity of peptides derived from a fibronectin-binding protein of S. aureus expressed on two different plant viruses (1999) Vaccine, 17, pp. 1846-1857; Chapman, S., Kavanagh, T., Baulcombe, D., Potato virus X as a vector for gene expression in plants (1992) Plant J, 2, pp. 549-557; Chapman, S., Hills, G., Watts, J., Baulcombe, D., Mutational analysis of the coat protein gene of potato virus X: Effects on virion morphology and viral pathogenicity (1992) Virology, 191, pp. 223-230; Chen, Y.-T., Scanlan, M.J., Sahin, U., A testicular antigen aberrantly expressed in human cancers detected by autologous antibody screening (1997) Proc Natl Acad Sci U S A, 94, pp. 1914-1918. , & 7 other authors; Donini, M., Lico, C., Baschieri, S., Conti, S., Magliani, W., Polonelli, L., Benvenuto, E., Production of an engineered killer peptide in Nicotiana benthamiana by using a Potato virus X expression system (2005) Appl Environ Microbiol, 71, pp. 6360-6367; Fedorkin, O.N., Solovyev, A.G., Yelina, N.E., Zamyatnin Jr., A.A., Zinovkin, R.A., Mikinen, K., Schiemann, J., Morozov, S.Yu., Cell-to-cell movement of potato virus X involves distinct functions of the coat protein (2001) J Gen Virol, 82, pp. 449-458; Hurwitz, J.L., Slobod, K.S., Lockey, T.D., Wang, S., Chou, T.-H.W., Lu, S., Application of the polyvalent approach to HIV-1 vaccine development (2005) Curr Drug Targets Infect Disord, 5, pp. 143-156; Kimalov, B., Gal-On, A., Stav, R., Belausov, E., Arazi, T., Maintenance of coat protein N-terminal net charge and not primary sequence is essential for zucchini yellow mosaic virus systemic infectivity (2004) J Gen Virol, 85, pp. 3421-3430; Kirkin, A.L., Dzhandzhugazyan, K., Zeuthen, J., Melanoma-associated antigens recognized by cytotoxic T lymphocytes (1998) APMIS, 106, pp. 665-679; Koenig, R., Torrance, L., Antigenic analysis of potato virus X by means of monoclonal antibodies (1986) J Gen Virol, 67, pp. 2145-2151; Kozlovsky, S.V., Karpova, O.V., Arkhipenko, M.V., Zayakina, O.V., Rodionova, N.P., Atabekov, I.G., Effect of the N-terminal domain of the coat protein of Potato virus X on the structure of viral particles (2003) Dokl Biochem Biophys, 391, pp. 189-191; Lee, A.G., Lipid-protein interactions in biological membranes: A structural perspective (2003) Biochim Biophys Acta, 1612, pp. 1-40; Lesemann, D.E., Cytopathology (1988) The Plant Viruses: The Filamentous Plant Viruses, pp. 179-235. , Edited by R. G. Milne. New York: Plenum; Lough, T.J., Shash, K., Xoconostle-Cizares, B., Hofstra, K.R., Beck, D.L., Balmori, E., Forster, R.L.S., Lucas, W.J., Molecular dissection of the mechanism by which potexvirus triple gene block proteins mediate cell-to-cell transport of infectious RNA (1998) Mol Plant Microbe Interact, 11, pp. 801-814; Lough, T.J., Netzler, N.E., Emerson, S.J., Sutherland, P., Carr, F., Beck, D.L., Lucas, W.J., Forster, R.L.S., Cell-to-cell movement of potexviruses: Evidence for a ribonucleoprotein complex involving the coat protein and the first triple gene block protein (2000) Mol Plant Microbe Interact, 13, pp. 962-974; Marusic, C., Rizza, P., Lattanzi, L., Mancini, C., Spada, M., Belardelli, F., Benvenuto, E., Capone, I., Chimeric plant virus particles as immunogens for inducing murine and human immune responses against human immunodeficiency virus type 1 (2001) J Virol, 75, pp. 8434-8439; Parker, L., Kendall, A., Stubbs, G., Surface features of potato virus X from fiber diffraction (2002) Virology, 300, pp. 291-295; Pogue, G.B., Lindbo, J.A., Garger, S.J., Fitzmaurice, W.P., Making an ally from an enemy: Plant virology and the new agriculture (2002) Annu Rev Phytopathol, 40, pp. 45-74; Polonelli, L., Magliani, W., Conti, S., Bracci, L., Lozzi, L., Neri, P., Adriani, D., Cassone, A., Therapeutic activity of an engineered synthetic killer antiidiotypic antibody fragment against experimental mucosal and systemic candidiasis (2003) Infect Immun, 71, pp. 6205-6212; Porta, C., Spall, V.E., Findlay, K.C., Gergerich, R.C., Farrance, C.E., Lomonossoff, G.P., Cowpea mosaic virus-based chimaeras. Effects of inserted peptides on the phenotype, host range, and transmissibility of the modified viruses (2003) Virology, 310, pp. 50-63; Salzwedel, K., West, J.T., Hunter, E., A conserved tryptophanrich motif in the membrane-proximal region of the human immunodeficiency virus type 1 gp41 ectodomain is important for Env-mediated fusion and virus infectivity (1999) J Virol, 73, pp. 2469-2480; Santa Cruz, S., Chapman, S., Roberts, A.G., Roberts, I.M., Prior, D.A.M., Oparka, K.J., Assembly and movement of a plant virus carrying a green fluorescent protein overcoat (1996) Proc Natl Acad Sci U S A, 93, pp. 6286-6290; Santa Cruz, S., Roberts, A.G., Prior, D.A.M., Chapman, S., Oparka, K.J., Cell-to-cell and phloem-mediated transport of potato virus X: The role of virions (1998) Plant Cell, 10, pp. 495-510; Scala, G., Chen, X., Liu, W., Telles, J.N., Cohen, O.J., Vaccarezza, M., Igarashi, T., Fauci, A.S., Selection of HIV-specific immunogenic epitopes by screening random peptide libraries with HIV-1-positive sera (1999) J Immunol, 162, pp. 6155-6161; Scholthof, H.B., Scholthof, K.-B.G., Jackson, A.O., Plant gene virus vectors for transient expression of foreign proteins in plants (1996) Annu Rev Phytopathol, 34, pp. 299-323; Shalla, T.A., Shepard, J.F., The structure and antigenic analysis of amorphous inclusion bodies induced by potato virus X (1972) Virology, 49, pp. 654-667; Shan, L., Molberg, øO., Parrot, I., Hausch, F., Filiz, F., Gray, G.M., Sollid, L.M., Khosla, C., Structural basis for gluten intolerance in celiac sprue (2002) Science, 297, pp. 2275-2279; Shanmugam, G., Polavarapu, P.L., Kendall, A., Stubbs, G., Structures of plant viruses from vibrational circular dichroism (2005) J Gen Virol, 86, pp. 2371-2377; Shiver, J.W., Emini, E.A., Recent advances in the development of HIV-1 vaccines using replication-incompetent adenovirus vectors (2004) Annu Rev Med, 55, pp. 355-372; Tangri, S., Mothé, B.R., Eisenbraun, J., Rationally engineered therapeutic proteins with reduced immunogenicity (2005) J Immunol, 174, pp. 3187-3196. , & 9 other authors