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. 1983 Oct;3(10):1746–1758. doi: 10.1128/mcb.3.10.1746

Rous sarcoma virus contains sequences which permit expression of the gag gene in Escherichia coli.

B Mermer, M Malamy, J M Coffin
PMCID: PMC370036  PMID: 6316124

Abstract

Several aspects of Rous sarcoma virus gene expression, including transcription, translation, and protein processing, can occur within Escherichia coli containing cloned viral DNA. The viral long terminal repeat contains a bacterial promoter, and viral sequences at or near the authentic viral initiation codon permit the initiation of translation. These signals can direct the synthesis in E. coli of the viral gag gene precursor Pr76 or, when fused to a portion of the lacZ gene, a gag-beta-galactosidase fusion protein. Pr76 is processed into gag structural proteins in E. coli in a process which is dependent upon the gag product p15. These observations suggest that E. coli can be used for the introduction and analysis of mutations in sequences relevant to viral gene expression.

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Selected References

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  1. Beckwith J. A genetic approach to characterizing complex promoters in E. coli. Cell. 1981 Feb;23(2):307–308. doi: 10.1016/0092-8674(81)90125-2. [DOI] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Bishop J. M., Courtneidge S. A., Levinson A. D., Oppermann H., Quintrell N., Sheiness D. K., Weiss S. R., Varmus H. E. Origin and function of avian retrovirus transforming genes. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):919–930. doi: 10.1101/sqb.1980.044.01.099. [DOI] [PubMed] [Google Scholar]
  4. Blair D. G., Oskarsson M., Wood T. G., McClements W. L., Fischinger P. J., Vande Woude G. G. Activation of the transforming potential of a normal cell sequence: a molecular model for oncogenesis. Science. 1981 May 22;212(4497):941–943. doi: 10.1126/science.7233190. [DOI] [PubMed] [Google Scholar]
  5. Blumberg D. D., Malamy M. H. Evidence for the presence of nontranslated T7 late mRNA in infected F'(PIF+) episome-containing cells. J Virol. 1974 Feb;13(2):378–385. doi: 10.1128/jvi.13.2.378-385.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bukhari A. I., Zipser D. Mutants of Escherichia coli with a defect in the degradation of nonsense fragments. Nat New Biol. 1973 Jun 20;243(129):238–241. doi: 10.1038/newbio243238a0. [DOI] [PubMed] [Google Scholar]
  7. Casadaban M. J., Chou J., Cohen S. N. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. doi: 10.1128/jb.143.2.971-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coffin J. M., Tsichlis P. N., Barker C. S., Voynow S., Robinson H. L. Variation in avian retrovirus genomes. Ann N Y Acad Sci. 1980;354:410–425. doi: 10.1111/j.1749-6632.1980.tb27982.x. [DOI] [PubMed] [Google Scholar]
  9. Conklin K. F., Coffin J. M., Robinson H. L., Groudine M., Eisenman R. Role of methylation in the induced and spontaneous expression of the avian endogenous virus ev-1: DNA structure and gene products. Mol Cell Biol. 1982 Jun;2(6):638–652. doi: 10.1128/mcb.2.6.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dittmar K. J., Moelling K. Biochemical properties of p15-associated protease in an avian RNA tumor virus. J Virol. 1978 Oct;28(1):106–118. doi: 10.1128/jvi.28.1.106-118.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eisenman R. N., Mason W. S., Linial M. Synthesis and processing of polymerase proteins of wild-type and mutant avian retroviruses. J Virol. 1980 Oct;36(1):62–78. doi: 10.1128/jvi.36.1.62-78.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guntaka R. V., Rao P. Y., Katz R. A., Mitsialis S. A. Binding of Escherichia coli RNA polymerase to a specific site located near the 3'-end of the avaian sarcoma virus genome. Biochim Biophys Acta. 1980 May 30;607(3):457–469. doi: 10.1016/0005-2787(80)90156-2. [DOI] [PubMed] [Google Scholar]
  13. Hsu T. W., Sabran J. L., Mark G. E., Guntaka R. V., Taylor J. M. Analysis of unintegrated avian RNA tumor virus double-stranded DNA intermediates. J Virol. 1978 Dec;28(3):810–818. doi: 10.1128/jvi.28.3.810-818.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Katz R. A., Omer C. A., Weis J. H., Mitsialis S. A., Faras A. J., Guntaka R. V. Restriction endonuclease and nucleotide sequence analyses of molecularly cloned unintegrated avian tumor virus DNA: structure of large terminal repeats in circle junctions. J Virol. 1982 Apr;42(1):346–351. doi: 10.1128/jvi.42.1.346-351.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leis J. P., McGinnis J., Green R. W. Rous sarcoma virus p19 binds to specific double-stranded regions of viral RNA: effect of p19 on cleavage of viral RNA by RNase III. Virology. 1978 Jan;84(1):87–98. doi: 10.1016/0042-6822(78)90220-9. [DOI] [PubMed] [Google Scholar]
  16. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  17. Mitsialis S. A., Young J. F., Palese P., Guntaka R. V. An avian tumor virus promoter directs expression of plasmid genes in Escherichia coli. Gene. 1981 Dec;16(1-3):217–225. doi: 10.1016/0378-1119(81)90078-0. [DOI] [PubMed] [Google Scholar]
  18. Moelling K., Scott A., Dittmar K. E., Owada M. Effect of p15-associated protease from an avian RNA tumor virus on avian virus-specific polyprotein precursors. J Virol. 1980 Feb;33(2):680–688. doi: 10.1128/jvi.33.2.680-688.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Neel B. G., Hayward W. S., Robinson H. L., Fang J., Astrin S. M. Avian leukosis virus-induced tumors have common proviral integration sites and synthesize discrete new RNAs: oncogenesis by promoter insertion. Cell. 1981 Feb;23(2):323–334. doi: 10.1016/0092-8674(81)90128-8. [DOI] [PubMed] [Google Scholar]
  20. Payne G. S., Courtneidge S. A., Crittenden L. B., Fadly A. M., Bishop J. M., Varmus H. E. Analysis of avian leukosis virus DNA and RNA in bursal tumours: viral gene expression is not required for maintenance of the tumor state. Cell. 1981 Feb;23(2):311–322. doi: 10.1016/0092-8674(81)90127-6. [DOI] [PubMed] [Google Scholar]
  21. Schwartz D. E., Tizard R., Gilbert W. Nucleotide sequence of Rous sarcoma virus. Cell. 1983 Mar;32(3):853–869. doi: 10.1016/0092-8674(83)90071-5. [DOI] [PubMed] [Google Scholar]
  22. Shank P. R., Hughes S. H., Kung H. J., Majors J. E., Quintrell N., Guntaka R. V., Bishop J. M., Varmus H. E. Mapping unintegrated avian sarcoma virus DNA: termini of linear DNA bear 300 nucleotides present once or twice in two species of circular DNA. Cell. 1978 Dec;15(4):1383–1395. doi: 10.1016/0092-8674(78)90063-6. [DOI] [PubMed] [Google Scholar]
  23. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Siebenlist U., Simpson R. B., Gilbert W. E. coli RNA polymerase interacts homologously with two different promoters. Cell. 1980 Jun;20(2):269–281. doi: 10.1016/0092-8674(80)90613-3. [DOI] [PubMed] [Google Scholar]
  25. Siegert W., Konings R. N., Bauer H., Hofschneider P. H. Translation of avian myeloblastosis virus RNA in a cell-free lysate of Escherichia coli. Proc Natl Acad Sci U S A. 1972 Apr;69(4):888–891. doi: 10.1073/pnas.69.4.888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Temin H. M., Baltimore D. RNA-directed DNA synthesis and RNA tumor viruses. Adv Virus Res. 1972;17:129–186. doi: 10.1016/s0065-3527(08)60749-6. [DOI] [PubMed] [Google Scholar]
  27. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tsichlis P. N., Coffin J. M. Recombinants between endogenous and exogenous avian tumor viruses: role of the C region and other portions of the genome in the control of replication and transformation. J Virol. 1980 Jan;33(1):238–249. doi: 10.1128/jvi.33.1.238-249.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tsichlis P. N., Coffin J. M. Role of the C region in relative growth rates of endogenous and exogenous avian oncoviruses. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):1123–1132. doi: 10.1101/sqb.1980.044.01.121. [DOI] [PubMed] [Google Scholar]
  30. Vogt V. M., Wight A., Eisenman R. In vitro cleavage of avian retrovirus gag proteins by viral protease p15. Virology. 1979 Oct 15;98(1):154–167. doi: 10.1016/0042-6822(79)90534-8. [DOI] [PubMed] [Google Scholar]
  31. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. von der Helm K., Kempeni J., Wille W., Willecke K. Rous sarcoma virus precursor protein pr 76 is processed in avian sarcoma virus-transformed mammalian cells after fusion-injection of viral protein p 15. Virology. 1980 Oct 30;106(2):310–316. doi: 10.1016/0042-6822(80)90254-8. [DOI] [PubMed] [Google Scholar]

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