Functional Env Research Articles

Helper virus-dependent RSV rescue from two lines of hamster cells transformed in vivo by XC RSV.

Newborn hamsters were inoculated with chicken sarcoma tissue produced by XC RSV which had been rescued by transfection from XC cells. Tumours with a latency of up to 20 months appeared in all inoculated hamsters. Three tumour cell lines were successfully established for in vitro growth and cloned. The virus was rescued from the PR RSH-9 cell line by cell fusion with chicken fibroblasts and from the other two lines (PR RSH-12 and PR RSH-14) and clones only if chicken fibroblasts used for fusion had been preinfected with a helper virus. The last two cell lines evidently harboured a functional env gene since the the synthesis of transforming virus started soon after they were found with 16Q cells producing defective BH-RSH(-) lacking the envelope glycoprotein. Produced viruses acquired subgroup C specificity of XC rsv originally used for transformation of these two cell lines. The nature of provirus alterations in PR RSH-12 and PR RSH-14 cells as well as the mechanisms underlying these changes are discussed.

Generation of nondefective Rous sarcoma virus by asymmetric recombination between deletion mutants.

A replication-defective deletion mutant of Prague Rous sarcoma virus (RSV), which lacks functional gag, pol, and env genes, was crossed with a transformation-defective deletion mutant derived from Schmidt-Ruppin RSV. Transformation- and replication-competent viruses were generated in the cross. Characterization of one of these rescued viruses indicated that it was a nondefective recombinant containing the src gene of the replication-defective mutant plus the replicative genes of the transformation-defective virus. These results indicate that, contrary to previous reports, asymmetric recombination between RSV deletion mutants can result in the formation of nondefective RSV.

A nonconditional replication-defective mutant of the Schmidt-Ruppin strain of Rous sarcoma virus

A nonconditional mutant of the Schmidt-Ruppin strain of Rous sarcoma virus (subgroup A) that fails to synthesize infectious progeny virus is described. Nonproducing cells transformed by this mutant LA7365 have functional viral src and env genes. The gag gene of LA7365 is defective. There is no complementation with a gag ts mutant, and pulse-chase experiments show that the proteolytic processing of the gag precursor pr76 is strongly inhibited in mutant-infected cells. Whether there is an additional lesion in the pol gene of LA 7365 could not be determined from the available data.

Mapping of biological functions on RNA of avian tumor viruses: location of regions required for transformation and determination of host range.

A map of the large T1 oligonucleotides of the RNA of Prague Rous sarcoma virus, strain B (Pr RSVb) has recently been established (Coffin and Billeter, submitted for publication). Since the RNA of Rous associated virus, type 1 (RAV-1) lacks many of the large 1 oligonucleotides of Pr RSV-B and contains others not present in the latter, the RNA of recombinants between RAV-1 and Pr RSV-B could be analyzed with regard to the origin of its sequences. Recombinants were selected for transforming capacity (characteristic for Pr RSV-B) and ability to grow on C/B chicken fibroblasts (characteristic for RAV-1). Four out of five recombinants examined had undergone at least two crossovers. The set of Pr RSV-B-specific oligonucleotides present in all recombinants defined an RNA region near the poly(A) segment; this must contain genetic information required for transformation required for transformation (the onc function). All recombinants lost a set of contiguous Pr RSV-B-specific oligonucleotides and concomitantly acquired a set of RAV-1-specific oligonucleotides. These define a region in the middle section of the oligonucleotide map, all or some of which must be required for determining growth capacity on C/B cells (the env function).