Characterization of recombinant polioviruses expressing regions of rotavirus VP4, hepatitis B surface antigen, and herpes simplex virus type 2 glycoprotein D

Abstract

Recombinant polioviruses expressing antigens from rotavirus, herpes simplex virus type 2, and hepatitis B virus were generated. Fusion of the heterologous polypeptides to the amino terminus of the poliovirus polyprotein did not prevent myristylation of VP0, suggesting a novel mechanism of myristylation for these recombinant viruses. The effects of the parental genetic background, different foreign sequences, and different insert sizes on growth characteristics were compared. Both the size and the nature of the heterologous sequence appeared to be factors influencing the growth and stability of recombinant polioviruses. All of the recombinants showed a temperature-sensitive phenotype, regardless of the genetic background (attenuated or wild type) from which they were derived. Preliminary studies with transgenic mice carrying the poliovirus receptor gene are discussed.

Release of virus-like particles from cells infected with poliovirus replicons which express human immunodeficiency virus type 1 Gag

“…This idea is consistent with previous studies which have found that amino acids other than the penultimate glycine are essential for the myristylation of a protein (18,63). Furthermore, in agreement with our results, Mattion et al have demonstrated myristylation of the poliovirus VP0 and VP4 proteins after the processing of foreign gene sequences positioned 5Ј to the P1 region (35). Previous studies established that expression of the HIV-1 Gag protein, in the absence of other HIV proteins, results in the intracellular expression of a 55-kDa protein which is transported to the plasma membrane, where immature virus parti-FIG.…”

“…In recent years, several groups have reported on attempts to develop poliovirus as an expression system that could ultimately be used as a vaccine vector (3,4,13,35,36,50,51). Poliovirus is attractive for use as a vector for several reasons.…”

“…Therefore, proteinase 2A must have the capacity to process at tyrosineglycine dipeptide bonds positioned within the context of foreign proteins. The cleavage of a 2A site within the context of a foreign protein in the poliovirus cDNA is unique, since other studies have described only cleavage at 3C sites (4,35).…”

Expression of Foreign Proteins by Poliovirus Polyprotein Fusion: Analysis of Genetic Stability Reveals Rapid Deletions and Formation of Cardioviruslike Open Reading Frames

“…is small (Ͻ400 nt) (35,36,52), a phenomenon seen also with dicistronic viruses (32). Although the fusion of small neutralization epitopes of rotavirus, herpes simplex virus type 2, and hepatitis B virus to PV polyprotein yielded genotypes of increased genetic stability, the resulting viral expression vectors were impaired in growth at 37°C, either because they expressed a temperature-sensitive phenotype or because they were retarded at the level of encapsidation (35,36,52). A detailed analysis of the genetic properties of these viruses after multiple rounds of replication has not been carried out.…”

“…Many attempts have been made to manipulate the genomes of RNA viruses so that they can function as expression vectors. Among these RNA viruses are Sindbis virus (8,48,51), Semliki Forest virus (31), influenza virus (19,30), tobacco etch virus (16), mengovirus (2,3), and poliovirus (PV) (1,6,9,10,12,14,22,23,32,35,36,38,44).…”