Expression of Human papillomavirus 16 E7ggg oncoprotein on N- and C-terminus of Potato virus X coat protein in bacterial and plant cells

Plchová, Helena; Moravec, Tomáš; Hoffmeisterová, Hana; Folwarczna, Jitka; Čeřovská, Noemi

doi:10.1016/j.pep.2011.01.008

Cited by 30 publications

(22 citation statements)

References 41 publications

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“…Our effort to express the E7 epitope in plants using a PVX vector is related to our previous research on the expression of mutated E6 and E7 oncoproteins of HPV 16 in plants (Plchova et al 2011;Cerovska et al 2008Cerovska et al , 2013. To test Fig.…”

Section: Resultsmentioning

confidence: 99%

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Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation

Vaculik

Plchová

Moravec

et al. 2014

Plant Cell Tiss Organ Cult

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Potato virus X (PVX) is widely used as a peptide presentation system in plant biotechnology, mostly by fusion of desired peptides to the N-terminus of its capsid protein (CP). Considering that some epitopes can interfere with the stability and/or self-assembly of PVX CP when fused to its N-terminus, we evaluated four other possible sites for fusion using the E7 epitope derived from human papillomavirus type 16 with different tags. We prepared eight different PVX CP constructs modified with the E7 epitope fused with the 6xHis tag in both orientations (6xHis-E7, E7-6xHis), cloned them into the PVX-based vector pGR106 and expressed them transiently in Nicotiana benthamina plants. Only the fusion site located after amino acid 23 led to systemic infection of plants and the production of recombinant proteins, but no viral particles were detected. When we replaced the 6xHis with StrepII tag, the modified virus infected plants systemically, expressed proteins assembled into viral particles and the epitopes were located on the particle surface. The results of this study indicate that the new position within the PVX CP can be used for peptide presentation on the surface of PVX particles and is promising for PVX based production of therapeutic compounds in plants.

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Section: Resultsmentioning

confidence: 99%

“…The presented antigens are mostly fused to the N-terminus of CP (O'Brien et al 2000;Uhde et al 2005;Massa et al 2008;Morgenfeld et al 2009). We have shown previously that the C-terminus of PVX CP can also be used for antigen display (Plchova et al 2011;Cerovska et al 2012Cerovska et al , 2013.…”

Section: Introductionmentioning

confidence: 99%

Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation

Vaculik

Plchová

Moravec

et al. 2014

Plant Cell Tiss Organ Cult

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“…[7][8][9] It should be noted that C-terminal PVX coat protein fusions of peptides and proteins have also been reported. [10,11] In addition to the genetic engineering capabilities, chemical modification enables additional functionalization post-harvest. We established bioconjugation protocols targeting surface-exposed lysines using N-hydroxysuccinimide or Cu(I)-catalyzed azide-alkyne cycloaddition (click chemistry) enabling efficient functionalization of PVX with biotin linkers, fluorophores, PEG, targeting ligands, etc.…”

Section: Introductionmentioning

confidence: 99%

“…Its special interest in biotechnology is due to three prominent properties: 1. Its structural stability over a broad range of pH levels (pH [2][3][4][5][6][7][8][9][10][11][12] and in the presence of various detergents; 2. The IgG-protein A complex interaction is reversible; while stable over a broad range of physiological conditions, dissociation is possible under controlled conditions (pH 3.5-4.5) without apparent loss of activity.…”

Section: Introductionmentioning

confidence: 99%

Production of Immunoabsorbent Nanoparticles by Displaying Single‐Domain Protein A on Potato Virus X

Uhde‐Holzem

McBurney

Tiu

et al. 2015

Macromolecular Bioscience

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The combination of antibodies with nanoparticles provides wide-ranging applications in biosensing. While several covalent presentation strategies have been established, there is need for alternative, non-covalent methods to provide a routine for scalable nanomanufacturing. We report the multivalent presentation of the B domain of Staphylococcus aureus protein A (SpAB) on potato virus X (PVX) nanoparticles. Three different synthetic strategies were used to obtain chimeric PVX(SpAB) filaments. The protein A fragments displayed on the surface of all three PVX chimeras remained fully functional as an immunoabsorbent for antibody capture enabling biosensing. The new biomaterials presented could find applications as diagnostic tools for biomedical or environmental monitoring.

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“…The PVX CP can be used for presentation by direct or indirect fusion of a desired protein or peptide to the N-terminus [10][11][12]. Recently, it has been also shown that the C-terminus of PVX CP can be used for protein presentation [13]. However, the Abstract: Potato virus X (PVX) is widely used as a peptide presentation system in plant biotechnology, mostly for transient expression of desired peptides fused to the N-terminus of its capsid protein (CP).…”

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confidence: 99%

New positions for peptide presentation in Potato virus X capsid protein

et al. 2015

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viruses represents potentially a promising new delivery and adjuvant system mainly for the following reasons: (i) the capsid of plant virus represents a scaffold exposing desired epitopes to the immune system in an abundant and organized manner and can serve as an adjuvant by itself, (ii) plant viruses are not infectious to animals, lowering the safety risk associated with their deployment, and (iii) scalability [3,4].Human papillomavirus (HPV) infection is one of the most common sexually transmitted infections worldwide. Infection by high risk HPVs is causally associated with cervical cancer, which represents the second most common cancer among women (after breast cancer) [5]. The most common oncogenic HPV genotypes are 16 and 18, together causing approximately 70 % of all cervical cancers, while HPV 16 alone causing 50 % of all cervical cancers [6]. The E7 oncoprotein plays an important role in both malignant transformation and in the maintenance of the transformed state of HPV infected cells. This represents a promising target for immunotherapy because the E7 oncoprotein is expressed in all cervical tumors and in precancerous lesions [7]. The main known role of E7 oncoprotein in virus cycle is to prevent cell death by binding tumor-suppressor protein Rb (pRb), resulting in the dissociation of pRB from E2F-family transcription factors and the premature cell progression into the S phase of the cell cycle which leads to uncontrollable cell division [8].Potato virus X (PVX) belongs to the order Tymovirales, family Alphaflexiviridae and genus Potexvirus. It is a filamentous, positive single stranded RNA (+ssRNA) virus. Virus particles are about 515 nm long and have about 13 nm in diameter. The PVX genome is capped, has a poly(A) tail and encodes five open reading frames (viral replicase, three movement proteins belonging to the triple-gene block family and capsid protein -approximately 1 300 CP subunits per virion) [9]. The PVX CP can be used for presentation by direct or indirect fusion of a desired protein or peptide to the N-terminus [10][11][12]. Recently, it has been also shown that the C-terminus of PVX CP can be used for protein presentation [13]. However, the Abstract: Potato virus X (PVX) is widely used as a peptide presentation system in plant biotechnology, mostly for transient expression of desired peptides fused to the N-terminus of its capsid protein (CP). Here we describe testing/investigation of new positions for peptide presentation based on seven putative surface loops of PVX CP. We performed bacterial expression of fourteen different PVX CPs modified by the insertion of the epitope derived from the E7 oncoprotein (E7 epitope) of Human papillomavirus type 16 fused with His tag. The expression from vector pMPM-A4Ω in Escherichia coli MC1061 was performed to evaluate the capacity of the PVX CP platform to tolerate the insertion of E7 epitope fused with His tag in the seven putative surface loops. The immunological characteristics of expressed epitopes were assessed by Western blot using both anti-P...

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Expression of Human papillomavirus 16 E7ggg oncoprotein on N- and C-terminus of Potato virus X coat protein in bacterial and plant cells

Cited by 30 publications

References 41 publications

Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation

Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation

Production of Immunoabsorbent Nanoparticles by Displaying Single‐Domain Protein A on Potato Virus X

New positions for peptide presentation in Potato virus X capsid protein

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