INFECTION of fibroblasts by avian sarcoma virus (ASV) leads to neoplastic transformation of the host cell. Genetic analyses have implicated specific viral genes in the transforming process1–4, and recent results suggest that a single viral gene is responsible4. Normal chicken cells contain DNA homologous to part of the ASV genome5–8; moreover, embryonic fibroblasts from certain strains of chickens can produce low titres of infectious type C viruses either spontaneously9 or in response to various inducing agents10. None of the viruses obtained from normal chicken cells, however, can transform fibroblasts, and results with molecular hybridisation indicate that the nucleotide sequences responsible for transformation by ASV are not part of the genetic complement of the normal cell11. We demonstrate here that the DNA of normal chicken cells contains nucleotide sequences closely related to at least a portion of the transforming gene(s) of ASV; in addition, we have found that similar sequences are widely distributed among DNA of avian species and that they have diverged roughly according to phylogenetic distances among the species. Our data are relevant to current hypotheses of the origin of the genomes of RNA tumour viruses12 and the potential role of these genomes in oncogenesis13.

DNA related to the transforming gene(s) of avian sarcoma viruses is present in normal avian DNA

Efficient transcription of RNA into DNA by avian sarcoma virus polymerase

Publisher Summary The discovery of RNA-directed DNA synthesis in disrupted virions of RNA tumor viruses added strong support to the hypothesis that information transfer from RNA to DNA exists in biological system. The chapter discusses the properties of the endogenous reaction carried out by the virion DNA polymerase. To study the endogenous reaction disrupted, virions are incubated with substrates in the absence of any added template and synthesis of DNA is observed using the RNA present in the virions as template. The chapter also discusses the general implications of RNA-directed DNA synthesis in relation to tumor viruses, neoplastic cells, and normal cells. It is hypothesized that the leukovirus RNA-directed DNA polymerase activity is an integral part of the ribonucleoprotein core of the virions. The cores are synthesized in cells as precursor particles and then are incorporated into complete virions when the virions are assembled by budding at the cell surface. This core enzyme system contains not only the template-primer RNA, a DNA polymerase that can transfer information from RNA to double-stranded DNA, but ancillary enzymes, such as polynucleotide ligase and nucleases, which may aid in integrating the viral information with cellular DNA. This suggests that the virion enzyme activity is related to normal cellular DNA polymerases, and that there are homologies between the amino acid sequences of the viral enzyme and normal cellular enzymes. The relationship of RNA-directed DNA synthesis to neoplasia depends upon the relationship of RNA tumor viruses to neoplasia, which is supported by three general hypotheses: the provirus model, the oncogene model, and the protovirus model.

RNA-Directed DNA Synthesis and RNA Tumor Viruses

Analysis of avian leukosis virus DNA and RNA in bursal tumors: Viral gene expression is not required for maintenance of the tumor state

Along with other carcinogens of physical or chemical origin, viruses are known to be associated with and to cause tumors in a variety of experimental animals. RNA tumor viruses in particular are widespread in many species of animals, and frequently cause sarcomas or leukemia. The isolation of fowl sarcoma-leukemia virus in the early 1910s by a number of investigators marked the first successful identification of such tumor viruses (Ellermann and Bang, 1909; Rous, 1911; Fujinami and Inamoto, 1914). The cell-virus systems used for the studies of basic aspects of the pathogenesis of avian tumor viruses illustrate the progress in methodology for studying animal viruses in general. Early work in animal hosts was gradually replaced by a system using the chorioallantoic membrane of eggs (Keogh, 1938), then by tissue culture cells (Manaker and Groupe, 1956). The establishment of an assay system for Rous sarcoma virus (RSV) in tissue culture cells (Temin and Rubin, 1958) eventually led to an era of quantitative studies on the mechanism of cellular alteration by viruses.

Cell Transformation by RNA Tumor Viruses

Cells producing Rous sarcoma virus contain virus-specific ribonucleic acid (RNA) which can be identified by hybridization to single-stranded deoxyribonucleic acid (DNA) synthesized with RNA-directed DNA polymerase. Hybridization was detected by either fractionation on hydroxyapatite or hydrolysis with single strand-specific nucleases. Similar results were obtained with both procedures. The hybrids formed between enzymatically synthesized DNA and viral RNA have a high order of thermal stability, with only minor evidence of mismatched nucleotide sequences. Virus-specific RNA is present in both nuclei and cytoplasm of infected cells. This RNA is remarkably heterogeneous in size, including molecules which are probably restricted to the nucleus and which sediment in their native state more rapidly than the viral genome. The nature of the RNA found in cytoplasmic fractions varies from preparation to preparation, but heterogeneous RNA (ca. 4-50S), smaller than the viral genome, is always present in substantial amounts.

Virus-Specific Ribonucleic Acid in Cells Producing Rous Sarcoma Virus: Detection and Characterization

Identification of virus-specific RNA in cells infected with Rous sarcoma virus.

The deoxyribonucleic acid (DNA) polymerase(s) of Rous sarcoma virus synthesizes two principal products-single-stranded DNA in the form of a DNA:ribonucleic acid (RNA) hybrid and double-stranded DNA All of the single-stranded product and 50% of the double-stranded product can be hybridized to 70S viral RNA These results, in combination with data obtained by analysis of the kinetics of double-stranded DNA synthesis, indicate that the synthesis of double-stranded DNA is a sequel to the synthesis of single-stranded DNA and is dependent upon the latter for the provision of initial template

Jo-Ann Leong

Papers

Virus-Specific Ribonucleic Acid in Cells Producing Rous Sarcoma Virus: Detection and Characterization

Identification of virus-specific RNA in cells infected with Rous sarcoma virus.

Deoxyribonucleic Acid Polymerases of Rous Sarcoma Virus: Kinetics of Deoxyribonucleic Acid Synthesis and Specificity of the Products

Deoxyribonucleic AcidPolymerases ofRousSarcoma Virus: Kinetics ofDeoxyribonucleic AcidSynthesis andSpecificity oftheProducts