Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1987 Dec;61(12):3889–3895. doi: 10.1128/jvi.61.12.3889-3895.1987

Mutational analysis of the 3' open reading frames and the splice junction at nucleotide 3225 of bovine papillomavirus type 1.

P L Hermonat 1, P M Howley 1
PMCID: PMC256007  PMID: 2824822

Abstract

Functional analysis of the 3' open reading frames (ORFs) of bovine papillomavirus type 1 (BPV-1) has been complicated by the organization of that part of the genome. A region between nucleotides (nt) 3173 and 3551 contains three overlapping ORFs (E2, E3, and E4), as well as a 3' splice junction at nt 3225 which is used by many of the BPV-1 transcripts. To more clearly assign functions to specific ORFs in this region, single-base substitution mutations were generated which introduced translational termination codons into each of the three ORFs; a fourth mutation substituted an A with a C at nt 3223, altering the 3' splice junction consensus sequence from AG to CG. The E3- and the E4-specific mutants were wild type in their abilities to transform susceptible mouse C127 cells, to replicate as stable plasmids, and to trans-activate the E2 conditional enhancer. The E2-specific termination mutant was defective for plasmid replication, transformation, and trans-activation and could not be complemented for efficient transformation of a flat cell line which expressed the full-length E2 gene product. The splice junction mutant was defective for transformation of C127 cells and of a flat cell line expressing the full-length E2 gene product. These data extend previous analyses of the 3' ORFs and suggest that a spliced E2 product is involved in cellular transformation. The splice junction mutant could replicate as a stable plasmid, indicating that there is no absolute requirement in plasmid replication for a viral gene product expressed solely from an mRNA using the 3' splice junction at nt 3225.

Full text

PDF
3889

Images in this article

3892Image
on p.3892
3893Image
on p.3893
3894Image
on p.3894

Selected References

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

  1. Amtmann E., Sauer G. Bovine papilloma virus transcription: polyadenylated RNA species and assessment of the direction of transcription. J Virol. 1982 Jul;43(1):59–66. doi: 10.1128/jvi.43.1.59-66.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker C. C., Howley P. M. Differential promoter utilization by the bovine papillomavirus in transformed cells and productively infected wart tissues. EMBO J. 1987 Apr;6(4):1027–1035. doi: 10.1002/j.1460-2075.1987.tb04855.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen E. Y., Howley P. M., Levinson A. D., Seeburg P. H. The primary structure and genetic organization of the bovine papillomavirus type 1 genome. Nature. 1982 Oct 7;299(5883):529–534. doi: 10.1038/299529a0. [DOI] [PubMed] [Google Scholar]
  4. DiMaio D. Nonsense mutation in open reading frame E2 of bovine papillomavirus DNA. J Virol. 1986 Feb;57(2):475–480. doi: 10.1128/jvi.57.2.475-480.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Doorbar J., Campbell D., Grand R. J., Gallimore P. H. Identification of the human papilloma virus-1a E4 gene products. EMBO J. 1986 Feb;5(2):355–362. doi: 10.1002/j.1460-2075.1986.tb04219.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dvoretzky I., Shober R., Chattopadhyay S. K., Lowy D. R. A quantitative in vitro focus assay for bovine papilloma virus. Virology. 1980 Jun;103(2):369–375. doi: 10.1016/0042-6822(80)90195-6. [DOI] [PubMed] [Google Scholar]
  7. Engel L. W., Heilman C. A., Howley P. M. Transcriptional organization of bovine papillomavirus type 1. J Virol. 1983 Sep;47(3):516–528. doi: 10.1128/jvi.47.3.516-528.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gorman C. M., Howard B. H., Reeves R. Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. Nucleic Acids Res. 1983 Nov 11;11(21):7631–7648. doi: 10.1093/nar/11.21.7631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Groff D. E., Lancaster W. D. Genetic analysis of the 3' early region transformation and replication functions of bovine papillomavirus type 1. Virology. 1986 Apr 15;150(1):221–230. doi: 10.1016/0042-6822(86)90281-3. [DOI] [PubMed] [Google Scholar]
  10. Heilman C. A., Engel L., Lowy D. R., Howley P. M. Virus-specific transcription in bovine papillomavirus-transformed mouse cells. Virology. 1982 May;119(1):22–34. doi: 10.1016/0042-6822(82)90061-7. [DOI] [PubMed] [Google Scholar]
  11. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lambert P. F., Spalholz B. A., Howley P. M. A transcriptional repressor encoded by BPV-1 shares a common carboxy-terminal domain with the E2 transactivator. Cell. 1987 Jul 3;50(1):69–78. doi: 10.1016/0092-8674(87)90663-5. [DOI] [PubMed] [Google Scholar]
  13. Law M. F., Byrne J. C., Howley P. M. A stable bovine papillomavirus hybrid plasmid that expresses a dominant selective trait. Mol Cell Biol. 1983 Nov;3(11):2110–2115. doi: 10.1128/mcb.3.11.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Neary K., Horwitz B. H., DiMaio D. Mutational analysis of open reading frame E4 of bovine papillomavirus type 1. J Virol. 1987 Apr;61(4):1248–1252. doi: 10.1128/jvi.61.4.1248-1252.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rabson M. S., Yee C., Yang Y. C., Howley P. M. Bovine papillomavirus type 1 3' early region transformation and plasmid maintenance functions. J Virol. 1986 Nov;60(2):626–634. doi: 10.1128/jvi.60.2.626-634.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sarver N., Byrne J. C., Howley P. M. Transformation and replication in mouse cells of a bovine papillomavirus--pML2 plasmid vector that can be rescued in bacteria. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7147–7151. doi: 10.1073/pnas.79.23.7147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sarver N., Rabson M. S., Yang Y. C., Byrne J. C., Howley P. M. Localization and analysis of bovine papillomavirus type 1 transforming functions. J Virol. 1984 Nov;52(2):377–388. doi: 10.1128/jvi.52.2.377-388.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schiller J. T., Vass W. C., Lowy D. R. Identification of a second transforming region in bovine papillomavirus DNA. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7880–7884. doi: 10.1073/pnas.81.24.7880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schiller J. T., Vass W. C., Vousden K. H., Lowy D. R. E5 open reading frame of bovine papillomavirus type 1 encodes a transforming gene. J Virol. 1986 Jan;57(1):1–6. doi: 10.1128/jvi.57.1.1-6.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schlegel R., Wade-Glass M., Rabson M. S., Yang Y. C. The E5 transforming gene of bovine papillomavirus encodes a small, hydrophobic polypeptide. Science. 1986 Jul 25;233(4762):464–467. doi: 10.1126/science.3014660. [DOI] [PubMed] [Google Scholar]
  22. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  23. Spalholz B. A., Lambert P. F., Yee C. L., Howley P. M. Bovine papillomavirus transcriptional regulation: localization of the E2-responsive elements of the long control region. J Virol. 1987 Jul;61(7):2128–2137. doi: 10.1128/jvi.61.7.2128-2137.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Spalholz B. A., Yang Y. C., Howley P. M. Transactivation of a bovine papilloma virus transcriptional regulatory element by the E2 gene product. Cell. 1985 Aug;42(1):183–191. doi: 10.1016/s0092-8674(85)80114-8. [DOI] [PubMed] [Google Scholar]
  25. Stenlund A., Zabielski J., Ahola H., Moreno-Lopez J., Pettersson U. Messenger RNAs from the transforming region of bovine papilloma virus type I. J Mol Biol. 1985 Apr 20;182(4):541–554. doi: 10.1016/0022-2836(85)90240-2. [DOI] [PubMed] [Google Scholar]
  26. Yang Y. C., Okayama H., Howley P. M. Bovine papillomavirus contains multiple transforming genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1030–1034. doi: 10.1073/pnas.82.4.1030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yang Y. C., Spalholz B. A., Rabson M. S., Howley P. M. Dissociation of transforming and trans-activation functions for bovine papillomavirus type 1. Nature. 1985 Dec 12;318(6046):575–577. doi: 10.1038/318575a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES