Canine parvovirus (CPV) can productively infect canine and feline cell lines whereas feline panleukopenia virus (FPV) is restricted to the latter. The major determinants of tropism are two amino acids in the sequence shared by the capsid proteins, VP1 and VP2. We have shown that a rodent parvovirus-derived transducing genome, containing the luciferase reporter, can be packaged by VP1 and VP2 from separate helper sources. Canine A72 cells and feline CFK cells were transduced with recombinant virions generated using VP1 and VP2 combinations from CPV and FPV. Both VP1 and VP2 were necessary for production of transducing virions. Efficient transduction of A72 cells required VP2 of CPV. Therefore, the capsid determinants of tropism for CPV and FPV are in VP2, although a source of VP1 is also necessary to produce infectious particles. The results extend similar observations on the tropic determinants of different strains of minute virus of mice.The feline group of autonomous parvoviruses includes feline panleukopenia virus (FPV) and canine parvovirus (CPV), which have greater than 98 % sequence identity but a distinct host range (Truyen et al., 1992). In cell culture, CPV productively infects both canine and feline cell lines whereas FPV is restricted to feline cells. Studies with CPV-FPV recombinant genomes have shown that the canine host range is determined by the sequence between map units 59 and 73, within the overlapping coding sequence for VP1 and VP2 (Parrish & Carmichael, 1986). Site-directed mutagenesis has implicated amino acids 93 and 323 (numbered from the N terminus of VP2) as the major determinants of host range .We previously described the production of transducing recombinant virions derived from the rodent parvovirus LuIII, Author for correspondence : Ian Maxwell.Fax j1 303 315 8272. e-mail Ian.Maxwell!uchsc.edu with substitution of reporter genes for the viral coding sequences (Maxwell et al., 1993 a). The recombinant genome could be packaged by capsid proteins either from LuIII or from related parvoviruses, including CPV or FPV (Maxwell et al., 1993 b ;Spitzer et al., 1996). In particular, packaging of the recombinant with capsid proteins supplied by a CPV helper, but not by a FPV helper, conferred the ability to transduce canine A72 cells efficiently (Spitzer et al., 1996). By packaging a heterologous genome the results confirmed that the tropic determinants for CPV and FPV were specified by the capsid and direct involvement of the encoding DNA was excluded.The capsids of CPV and FPV are composed of 60 capsomers, which consist of VP1 (" 15 % of the mass of the capsid) and VP2 (Tsao et al., 1991 ; Agbandje et al., 1993). These proteins are translated from alternately spliced transcripts from the P38 promoter. However, since the amino acid residues that determine tropism are present in both VP1 and VP2, this function might be mediated by either protein.Knowledge of which protein is involved would help to clarify their roles in establishing infection. Here, we have investigated this question using th...
The autonomous parvoviruses are small, non-enveloped, single strand DNA viruses. They occur in many species and they have oncolytic properties. We are modifying the capsid of feline panleukopenia virus (FPV), a parvovirus which normally infects feline cells, with the goal of targeting human tumor cells for potential cancer therapy. Using recombinant viruses transducing a luciferase reporter, we show that insertion of a cyclically constrained, integrin-binding peptide at an exposed position on the FPV capsid enables transduction of an ␣v integrin-expressing human rhabdomyosarcoma cell line (Rh18A). These cells were not transduced by virus
The minute virus of mice, prototype strain MVMp, productively infects cultured murine fibroblasts but not T cells. The immunosuppressive strain, MVMi, shows the converse tropism. These reciprocal tropisms are mediated by the viral capsids, in which their determinants have been mapped to a few specific amino acids in the primary sequence shared by VP1 and VP2. Which of these proteins is relevant in presenting these determinants during infection is not known. We have approached this question using a recombinant parvovirus system in which a LuIII-derived transducing genome, containing the luciferase reporter in place of viral coding sequences, can be packaged by capsid proteins from separate helper sources. We generated transducing virions by using helper constructs expressing either VP1 or VP2, containing the MVMp or MVMi tropic determinant region, in various combinations. The virions were used to infect human NB324K cells and murine A9 fibroblasts. Transduction of the human cells (permissive for both MVMp and MVMi) required both VP1 and VP2 and was successful with all combinations of these proteins. In contrast, significant transducing activity for A9 cells was detected only with recombinant virions containing VP2 of MVMp, while the use of either source of VP1 had little effect. We conclude that VP2 from MVMp is necessary to enable infection of murine A9 fibroblasts.
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