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. 1979 Nov;32(2):567–582. doi: 10.1128/jvi.32.2.567-582.1979

Deletion mutant of the Bratislava-77 strain of Rous sarcoma virus containing a fusion of the group-specific antigen and envelope genes.

P M Dierks, P E Highfield, J T Parsons
PMCID: PMC353589  PMID: 91686

Abstract

The genetic compositions of two independently derived preparations of the Bratislava-77 strain (B77) of Rous sarcoma virus were analyzed after each was passaged seven or more times in duck embryo fibroblasts. RNase, T1-resistant oligonucleotide fingerprint analysis of virion RNA from both preparations of duck-passaged B77 revealed the presence of two large noncontiguous deletions. Approximately 75% of the RNAs contained a deletion which spans oligonucleotides 304 to 4 on the viral genome (about 3,500 nucleotides) and encompasses all of the B77 polymerase gene. More than 90% of the RNAs also contained a deletion which spans src-specific oligonucleotides 6 and 5(about 2,200 nucleotides) and is identical to the deletion observed in transformation-defective B77. Virion RNA from duck-passaged B77 also contained two oligonucleotides (D1 and D2) not observed in the RNA of B77 virus grown on chicken embryo fibroblasts. Analysis of the virion RNA of duck-passaged B77 by denaturing agarose gel electrophoresis revealed four major subunits with molecular weights of 3.40 x 10(6), 2.65 x 10(6), 2.25 x 10(6), and 1.55 x 10(6). Whereas the 3.40- and 2.65-megadalton (Mdal) RNA species comigrated with the nondefective and transformation-defective RNAs of B77 propagated on chicken embryo fibroblasts, no counterparts to the 2.25- and 1.55-Mdal RNAs were observed in the RNA of B77 grown on chicken embryo fibroblasts. Oligonucleotide fingerprint analysis of these RNA species revealed that the 2.65-Mdal RNA contains the src-specific deletion and that 2.25-Mdal RNA contains the polymerase region deletion; both of these deletions were observed in the 1.55-Mdal RNA, which was the major RNA subunit species detected in duck-passaged B77. The new oligonucleotides (D1 and D2) observed in the duck-passaged virus were present in the 2.25- and 1.55-Mdal RNA species in vitro and in vivo and directs the synthesis of a 130,000-dalton protein (p130). p130 contains antigenic determinants specific for p27 (gag gene) and gp85 (env gene) but does not contain sequences which cross-react with antisera directed against the alpha beta form of RNA-dependent DNA polymerase (pol gene). This RNA, therefore, is generated by a fusion of the gag and env genes of Rous sarcoma virus B77.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams J. M., Jeppesen P. G., Sanger F., Barrell B. G. Nucleotide sequence from the coat protein cistron of R17 bacteriophage RNA. Nature. 1969 Sep 6;223(5210):1009–1014. doi: 10.1038/2231009a0. [DOI] [PubMed] [Google Scholar]
  2. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  3. Beemon K., Hunter T. In vitro translation yields a possible Rous sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3302–3306. doi: 10.1073/pnas.74.8.3302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Billeter M. A., Parsons J. T., Coffin J. M. The nucleotide sequence complexity of avian tumor virus RNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3560–3564. doi: 10.1073/pnas.71.9.3560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  6. Cashion L. M., Joho R. H., Planitz M. A., Billeter M. A., Weissmann C. Initiation sites of Rous sarcoma virus RNA-directed DNA synthesis in vitro. Nature. 1976 Jul 15;262(5565):186–190. doi: 10.1038/262186a0. [DOI] [PubMed] [Google Scholar]
  7. Coffin J. M., Billeter M. A. A physical map of the Rous sarcoma virus genome. J Mol Biol. 1976 Jan 25;100(3):293–318. doi: 10.1016/s0022-2836(76)80065-4. [DOI] [PubMed] [Google Scholar]
  8. Coffin J. M., Champion M., Chabot F. Nucleotide sequence relationships between the genomes of an endogenous and an exogenous avian tumor virus. J Virol. 1978 Dec;28(3):972–991. doi: 10.1128/jvi.28.3.972-991.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Diggelmann H. Biosynthesis of an unglycosylated envelope glycoprotein of Rous sarcoma virus in the presence of tunicamycin. J Virol. 1979 Jun;30(3):799–804. doi: 10.1128/jvi.30.3.799-804.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Duesberg P. H., Kawai S., Wang L. H., Vogt P. K., Murphy H. M., Hanafusa H. RNA of replication-defective strains of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1569–1573. doi: 10.1073/pnas.72.4.1569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eisenman R., Shaikh R., Mason W. S. Identification of an avian oncovirus polyprotein in uninfected chick cells. Cell. 1978 May;14(1):89–104. doi: 10.1016/0092-8674(78)90304-5. [DOI] [PubMed] [Google Scholar]
  13. Gupta R. C., Randerath K. Use of specific endonuclease cleavage in RNA sequencing-an enzymic method for distinguishing between cytidine and uridine residues. Nucleic Acids Res. 1977 Oct;4(10):3441–3454. doi: 10.1093/nar/4.10.3441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. HANAFUSA H., HANAFUSA T., RUBIN H. ANALYSIS OF THE DEFECTIVENESS OF ROUS SARCOMA VIRUS, II. SPECIFICATION OF RSV ANTIGENICITY BY HELPER VIRUS. Proc Natl Acad Sci U S A. 1964 Jan;51:41–48. doi: 10.1073/pnas.51.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Halpern M. S., Bolognesi D. P., Lewandowski L. J. Isolation of the major viral glycoprotein and a putative precursor from cells transformed by avian sarcoma viruses. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2342–2346. doi: 10.1073/pnas.71.6.2342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hanafusa H., Hanafusa T. Noninfectious RSV deficient in DNA polymerase. Virology. 1971 Jan;43(1):313–316. doi: 10.1016/0042-6822(71)90251-0. [DOI] [PubMed] [Google Scholar]
  17. Hayward W. S. Size and genetic content of viral RNAs in avian oncovirus-infected cells. J Virol. 1977 Oct;24(1):47–63. doi: 10.1128/jvi.24.1.47-63.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Joho R. H., Billeter M. A., Weissmann C. Mapping of biological functions on RNA of avian tumor viruses: location of regions required for transformation and determination of host range. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4772–4776. doi: 10.1073/pnas.72.12.4772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kawai S., Hanafusa H. Isolation of defective mutant of avian sarcoma virus. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3493–3497. doi: 10.1073/pnas.70.12.3493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Krzyzek R. A., Collett M. S., Lau A. F., Perdue M. L., Leis J. P., Faras A. J. Evidence for splicing of avian sarcoma virus 5'-terminal genomic sequences into viral-specific RNA in infected cells. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1284–1288. doi: 10.1073/pnas.75.3.1284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Lewandowski L. J., Smith R. E., Bolognesi D. P., Halpern M. S. Viral glycoprotein synthesis under conditions of glucosamine block in cells transformed by avian sarcoma viruses. Virology. 1975 Aug;66(2):347–355. doi: 10.1016/0042-6822(75)90208-1. [DOI] [PubMed] [Google Scholar]
  23. Linial M., Brown S., Neiman P. A nonconditional mutant of Rous sarcoma virus containing defective polymerase. Virology. 1978 Jun 1;87(1):130–141. doi: 10.1016/0042-6822(78)90165-4. [DOI] [PubMed] [Google Scholar]
  24. Linial M., Mason W. S. Characterization of two conditional early mutants of Rous sarcoma virus. Virology. 1973 May;53(1):258–273. doi: 10.1016/0042-6822(73)90484-4. [DOI] [PubMed] [Google Scholar]
  25. Linial M., Medeiros E., Hayward W. S. An avian oncovirus mutant (SE 21Q1b) deficient in genomic RNA: biological and biochemical characterization. Cell. 1978 Dec;15(4):1371–1381. doi: 10.1016/0092-8674(78)90062-4. [DOI] [PubMed] [Google Scholar]
  26. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mellon P., Duesberg P. H. Subgenomic, cellular Rous sarcoma virus RNAs contain oligonucleotides from the 3' half and the 5' terminus of virion RNA. Nature. 1977 Dec 15;270(5638):631–634. doi: 10.1038/270631a0. [DOI] [PubMed] [Google Scholar]
  28. Moelling K., Hayami M. Analysis of precursors to the envelope glycoproteins of avian RNA tumor viruses in chicken and quail cells. J Virol. 1977 Jun;22(3):598–607. doi: 10.1128/jvi.22.3.598-607.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Parsons J. T., Lewis P., Dierks P. Purification of virus-specific RNA from chicken cells infected with avian sarcoma virus: identification of genome-length and subgenome-leghth viral RNAs. J Virol. 1978 Jul;27(1):227–238. doi: 10.1128/jvi.27.1.227-238.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  31. Purchio A. F., Erikson E., Erikson R. L. Translation of 35S and of subgenomic regions of avian sarcoma virus RNA. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4661–4665. doi: 10.1073/pnas.74.10.4661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shine J., Czernilofsky A. P., Friedrich R., Bishop J. M., Goodman H. M. Nucleotide sequence at the 5' terminus of the avian sarcoma virus genome. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1473–1477. doi: 10.1073/pnas.74.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stoll E., Billeter M. A., Palmenberg A., Weissmann C. Avian myeloblastosis virus RNA is terminally redundant: implications for the mechanism of retrovirus replication. Cell. 1977 Sep;12(1):57–72. doi: 10.1016/0092-8674(77)90185-4. [DOI] [PubMed] [Google Scholar]
  34. TEMIN H. M. Separation of morphological conversion and virus production in Rous sarcoma virus infection. Cold Spring Harb Symp Quant Biol. 1962;27:407–414. doi: 10.1101/sqb.1962.027.001.038. [DOI] [PubMed] [Google Scholar]
  35. Vogt P. K. Spontaneous segregation of nontransforming viruses from cloned sarcoma viruses. Virology. 1971 Dec;46(3):939–946. doi: 10.1016/0042-6822(71)90092-4. [DOI] [PubMed] [Google Scholar]
  36. Wang L. H., Duesberg P. H., Kawai S., Hanafusa H. Location of envelope-specific and sarcoma-specific oligonucleotides on RNA of Schmidt-Ruppin Rous sarcoma virus. Proc Natl Acad Sci U S A. 1976 Feb;73(2):447–451. doi: 10.1073/pnas.73.2.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wang L. H., Duesberg P., Beemon K., Vogt P. K. Mapping RNase T1-resistant oligonucleotides of avian tumor virus RNAs: sarcoma-specific oligonucleotides are near the poly(A) end and oligonucleotides common to sarcoma and transformation-defective viruses are at the poly(A) end. J Virol. 1975 Oct;16(4):1051–1070. doi: 10.1128/jvi.16.4.1051-1070.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wang S. Y., Hayward W. S., Hanafusa H. Genetic variation in the RNA transcripts of endogenous virus genes in uninfected chicken cells. J Virol. 1977 Oct;24(1):64–73. doi: 10.1128/jvi.24.1.64-73.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Weiss R. A., Mason W. S., Vogt P. K. Genetic recombinants and heterozygotes derived from endogenous and exogenous avian RNA tumor viruses. Virology. 1973 Apr;52(2):535–552. doi: 10.1016/0042-6822(73)90349-8. [DOI] [PubMed] [Google Scholar]
  40. Weiss S. R., Varmus H. E., Bishop J. M. The size and genetic composition of virus-specific RNAs in the cytoplasm of cells producing avian sarcoma-leukosis viruses. Cell. 1977 Dec;12(4):983–992. doi: 10.1016/0092-8674(77)90163-5. [DOI] [PubMed] [Google Scholar]

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