Peptide display on Potato virus X: molecular features of the coat protein-fused peptide affecting cell-to-cell and phloem movement of chimeric virus particles.
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 peptides 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.
Lico
Chiara
C
Sezione di Genetica e Genomica Vegetale, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, Italy.
Capuano
Floriana
F
Sezione di Genetica e Genomica Vegetale, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, Italy.
Renzone
Giovanni
G
Proteomics and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147 Naples, Italy.
Donini
Marcello
M
Sezione di Genetica e Genomica Vegetale, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, Italy.
Marusic
Carla
C
Sezione di Genetica e Genomica Vegetale, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, Italy.
Scaloni
Andrea
A
Proteomics and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147 Naples, Italy.
Benvenuto
Eugenio
E
Sezione di Genetica e Genomica Vegetale, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, Italy.
Baschieri
Selene
S
Sezione di Genetica e Genomica Vegetale, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, Italy.
D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D009068:Movement; D009154:Mutation; D018515:Plant Leaves / Q000166:cytology / Q000648:ultrastructure / Q000821:virology*; D010957:Plasmids; D017863:Potexvirus / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology* / Q000821:virology*
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins