Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1992 Dec;66(12):7153–7158. doi: 10.1128/jvi.66.12.7153-7158.1992

Mutations in the VP1 coding region of polyomavirus determine differentiating stage specificity.

L Ricci 1, R Maione 1, C Passananti 1, A Felsani 1, P Amati 1
PMCID: PMC240404  PMID: 1331516

Abstract

Polyomavirus mutants capable of replicating in undifferentiated murine C2 myoblasts were selected and characterized. These mutants grow normally in 3T6 mouse fibroblast cells, and they do not complement the wild-type virus in coinfection experiments of C2 myoblasts. Of 12 isolates, 10 possess duplications of the regulatory region including the enhancer A domain. On the bases of the regulatory region structure and the presence and length of the enhancer duplication, the mutant viruses could be grouped into three classes. One mutant class (e.g., PyMB3) possesses an enhancer duplication of 91 bp identical to that of a previously characterized polyomavirus mutant, PyNB11/1. We have demonstrated that this enhancer duplication gives rise at its junction to a novel recognition motif for the transcriptional factor NF-1 (M. Caruso, C. Iacobini, C. Passananti, A. Felsani, and P. Amati, EMBO J. 9:947-955, 1990). The regulatory region PyMB3 virus recombined in a wild-type genome context maintains the mutant phenotype. The other two types of mutants, one with a 30-bp enhancer duplication (e.g., PyMB40) and one with a wild-type enhancer structure (e.g., PyMB27), possess two similar but distinct 6-bp deletions in the same region of the VP1 coding gene. In both cases, the ability to replicate in undifferentiated C2 myoblasts is strictly correlated to the mutation in the VP1 coding region.

Full text

PDF
7153

Images in this article

Selected References

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

  1. Amati P. Polyoma regulatory region: a potential probe for mouse cell differentiation. Cell. 1985 Dec;43(3 Pt 2):561–562. doi: 10.1016/0092-8674(85)90225-9. [DOI] [PubMed] [Google Scholar]
  2. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buckler-White A. J., Humphrey G. W., Pigiet V. Association of polyoma T antigen and DNA with the nuclear matrix from lytically infected 3T6 cells. Cell. 1980 Nov;22(1 Pt 1):37–46. doi: 10.1016/0092-8674(80)90152-x. [DOI] [PubMed] [Google Scholar]
  4. Caruso M., Iacobini C., Passananti C., Felsani A., Amati P. Protein recognition sites in polyomavirus enhancer: formation of a novel site for NF-1 factor in an enhancer mutant and characterization of a site in the enhancer D domain. EMBO J. 1990 Mar;9(3):947–955. doi: 10.1002/j.1460-2075.1990.tb08193.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. De Simone V., Amati P. Replicative cis-advantage of polyomavirus regulatory region mutants in different murine cell lines. J Virol. 1987 May;61(5):1615–1620. doi: 10.1128/jvi.61.5.1615-1620.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dubensky T. W., Freund R., Dawe C. J., Benjamin T. L. Polyomavirus replication in mice: influences of VP1 type and route of inoculation. J Virol. 1991 Jan;65(1):342–349. doi: 10.1128/jvi.65.1.342-349.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dynan W. S. Modularity in promoters and enhancers. Cell. 1989 Jul 14;58(1):1–4. doi: 10.1016/0092-8674(89)90393-0. [DOI] [PubMed] [Google Scholar]
  8. Felsani A., Maione R., Ricci L., Amati P. Coordinate expression of myogenic functions and polyoma virus replication. Cold Spring Harb Symp Quant Biol. 1985;50:753–757. doi: 10.1101/sqb.1985.050.01.093. [DOI] [PubMed] [Google Scholar]
  9. Freund R., Calderone A., Dawe C. J., Benjamin T. L. Polyomavirus tumor induction in mice: effects of polymorphisms of VP1 and large T antigen. J Virol. 1991 Jan;65(1):335–341. doi: 10.1128/jvi.65.1.335-341.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Freund R., Garcea R. L., Sahli R., Benjamin T. L. A single-amino-acid substitution in polyomavirus VP1 correlates with plaque size and hemagglutination behavior. J Virol. 1991 Jan;65(1):350–355. doi: 10.1128/jvi.65.1.350-355.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fujimura F. K., Deininger P. L., Friedmann T., Linney E. Mutation near the polyoma DNA replication origin permits productive infection of F9 embryonal carcinoma cells. Cell. 1981 Mar;23(3):809–814. doi: 10.1016/0092-8674(81)90445-1. [DOI] [PubMed] [Google Scholar]
  12. Glenn G. M., Eckhart W. Transcriptional regulation of early-response genes during polyomavirus infection. J Virol. 1990 May;64(5):2193–2201. doi: 10.1128/jvi.64.5.2193-2201.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  14. Jones N. C., Rigby P. W., Ziff E. B. Trans-acting protein factors and the regulation of eukaryotic transcription: lessons from studies on DNA tumor viruses. Genes Dev. 1988 Mar;2(3):267–281. doi: 10.1101/gad.2.3.267. [DOI] [PubMed] [Google Scholar]
  15. Katinka M., Vasseur M., Montreau N., Yaniv M., Blangy D. Polyoma DNA sequences involved in control of viral gene expression in murine embryonal carcinoma cells. Nature. 1981 Apr 23;290(5808):720–722. doi: 10.1038/290720a0. [DOI] [PubMed] [Google Scholar]
  16. Liddington R. C., Yan Y., Moulai J., Sahli R., Benjamin T. L., Harrison S. C. Structure of simian virus 40 at 3.8-A resolution. Nature. 1991 Nov 28;354(6351):278–284. doi: 10.1038/354278a0. [DOI] [PubMed] [Google Scholar]
  17. Lusky M., Berg L., Weiher H., Botchan M. Bovine papilloma virus contains an activator of gene expression at the distal end of the early transcription unit. Mol Cell Biol. 1983 Jun;3(6):1108–1122. doi: 10.1128/mcb.3.6.1108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Maione R., Felsani A., Pozzi L., Caruso M., Amati P. Polyomavirus genome and polyomavirus enhancer-driven gene expression during myogenesis. J Virol. 1989 Nov;63(11):4890–4897. doi: 10.1128/jvi.63.11.4890-4897.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Maione R., Passananti C., De Simone V., Delli-Bovi P., Augusti-Tocco G., Amati P. Selection of mouse neuroblastoma cell-specific polyoma virus mutants with stage differentiative advantages of replication. EMBO J. 1985 Dec 1;4(12):3215–3221. doi: 10.1002/j.1460-2075.1985.tb04068.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McCutchan J. H., Pagano J. S. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J Natl Cancer Inst. 1968 Aug;41(2):351–357. [PubMed] [Google Scholar]
  21. Melucci-Vigo G., Ciotta C., Risuleo G. cis-acting sequences that control the level of viral DNA synthesis in the polyomavirus late region. J Virol. 1989 Nov;63(11):4979–4981. doi: 10.1128/jvi.63.11.4979-4981.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  23. Moreland R. B., Montross L., Garcea R. L. Characterization of the DNA-binding properties of the polyomavirus capsid protein VP1. J Virol. 1991 Mar;65(3):1168–1176. doi: 10.1128/jvi.65.3.1168-1176.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rochford R., Campbell B. A., Villarreal L. P. Genetic analysis of the enhancer requirements for polyomavirus DNA replication in mice. J Virol. 1990 Feb;64(2):476–485. doi: 10.1128/jvi.64.2.476-485.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Santoro C., Mermod N., Andrews P. C., Tjian R. A family of human CCAAT-box-binding proteins active in transcription and DNA replication: cloning and expression of multiple cDNAs. Nature. 1988 Jul 21;334(6179):218–224. doi: 10.1038/334218a0. [DOI] [PubMed] [Google Scholar]
  26. Sekikawa K., Levine A. J. Isolation and characterization of polyoma host range mutants that replicate in nullipotential embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1100–1104. doi: 10.1073/pnas.78.2.1100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stamatos N. M., Chakrabarti S., Moss B., Hare J. D. Expression of polyomavirus virion proteins by a vaccinia virus vector: association of VP1 and VP2 with the nuclear framework. J Virol. 1987 Feb;61(2):516–525. doi: 10.1128/jvi.61.2.516-525.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tseng R. W., Fujimura F. K. Multiple domains in the polyomavirus B enhancer are required for productive infection of F9 embryonal carcinoma cells. J Virol. 1988 Aug;62(8):2890–2895. doi: 10.1128/jvi.62.8.2890-2895.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tseng R. W., Williams T., Fujimura F. K. Unique requirement for the PyF441 mutation for polyomavirus infection of F9 embryonal carcinoma cells. J Virol. 1988 Aug;62(8):2896–2902. doi: 10.1128/jvi.62.8.2896-2902.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
  31. Veldman G. M., Lupton S., Kamen R. Polyomavirus enhancer contains multiple redundant sequence elements that activate both DNA replication and gene expression. Mol Cell Biol. 1985 Apr;5(4):649–658. doi: 10.1128/mcb.5.4.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Villarreal L. P. Relationship of eukaryotic DNA replication to committed gene expression: general theory for gene control. Microbiol Rev. 1991 Sep;55(3):512–542. doi: 10.1128/mr.55.3.512-542.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yaffe D., Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature. 1977 Dec 22;270(5639):725–727. doi: 10.1038/270725a0. [DOI] [PubMed] [Google Scholar]
  34. Zullo J., Stiles C. D., Garcea R. L. Regulation of c-myc and c-fos mRNA levels by polyomavirus: distinct roles for the capsid protein VP1 and the viral early proteins. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1210–1214. doi: 10.1073/pnas.84.5.1210. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES