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. 1996 Nov;70(11):8003–8009. doi: 10.1128/jvi.70.11.8003-8009.1996

Insufficient levels of NFIII and its low affinity for the origin of adenovirus type 12 (Ad12) DNA replication contribute to the abortive infection of BHK21 hamster cells by Ad12.

G Schiedner 1, W Doerfler 1
PMCID: PMC190873  PMID: 8892924

Abstract

Human adenovirus type 12 (Ad12) induces undifferentiated sarcomas in neonate Syrian hamsters and hence presents a suitable model for studies of the molecular mechanism of viral oncogenesis. Since we submit that an understanding of the early steps in the interaction between Ad12 and hamster cells might shed light on the initiation of malignant transformation, the abortive infection of BHK21 hamster cells with Ad12 has been investigated in detail. Ad12 replication in these cells is blocked in early stages, while Ad2 can replicate to moderate titers. Early Ad12 genes are expressed in BHK21 hamster cells, but there is a total block in Ad12 DNA replication and late gene transcription. The Ad5-transformed hamster cell line BHK297-C131, with the left terminus of Ad5 DNA chromosomally integrated and constitutively expressed, allows limited levels of Ad12 DNA replication and late transcription, probably through Ad5 E1 functions, but not the translation of late Ad12 gene products. We have now investigated the capacities of binding of nuclear proteins NFI and NFIII from permissive human KB cells, nonpermissive hamster BHK21 cells, and complementing BHK297-C131 cells to the origin of replication (ori) of Ad2 or Ad12 DNA. The electrophoretic mobility shift assay has been used to assess these binding reactions. The data support the notions that NFIII of BHK21 cells has a lower affinity for the ori of Ad12 DNA than for the ori of Ad2 DNA and that the levels of NFIII in BHK21 cells are markedly reduced compared with the levels in the permissive human KB cells or the complementing BHK297-C131 hamster cells. These deficiencies are contributing factors for the abortive infection of BHK21 hamster cells with Ad12. The lack of sufficient levels of NFIII in BHK21 cells is also consistent with the decreased replication capacity of Ad2 in hamster compared with human cell lines.

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Selected References

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  1. BABLANIAN R., EGGERS H. J., TAMM I. STUDIES ON THE MECHANISM OF POLIOVIRUS-INDUCED CELL DAMAGE. I. THE RELATION BETWEEN POLIOVIRUS,-INDUCED METABOLIC AND MORPHOLOGICAL ALTERATIONS IN CULTURED CELLS. Virology. 1965 May;26:100–113. doi: 10.1016/0042-6822(65)90030-9. [DOI] [PubMed] [Google Scholar]
  2. Challberg M. D., Kelly T. J., Jr Adenovirus DNA replication in vitro. Proc Natl Acad Sci U S A. 1979 Feb;76(2):655–659. doi: 10.1073/pnas.76.2.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Challberg M. D., Rawlins D. R. Template requirements for the initiation of adenovirus DNA replication. Proc Natl Acad Sci U S A. 1984 Jan;81(1):100–104. doi: 10.1073/pnas.81.1.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chowrira B. M., Lucher L. A. Extracts of hamster cells abortively infected with human adenovirus type 12 are competent to support initiation of viral DNA replication. Virology. 1990 May;176(1):289–291. doi: 10.1016/0042-6822(90)90255-p. [DOI] [PubMed] [Google Scholar]
  5. Chowrira B. M., Zhao J. S., Lucher L. A. Formation in vitro of the pTP-dCMP initiation complex of human adenovirus type 12. J Gen Virol. 1991 Feb;72(Pt 2):427–430. doi: 10.1099/0022-1317-72-2-427. [DOI] [PubMed] [Google Scholar]
  6. Coenjaerts F. E., De Vries E., Pruijn G. J., Van Driel W., Bloemers S. M., Van der Lugt N. M., Van der Vliet P. C. Enhancement of DNA replication by transcription factors NFI and NFIII/Oct-1 depends critically on the positions of their binding sites in the adenovirus origin of replication. Biochim Biophys Acta. 1991 Aug 27;1090(1):61–69. doi: 10.1016/0167-4781(91)90037-m. [DOI] [PubMed] [Google Scholar]
  7. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Doerfler W. Nonproductive infection of baby hamster kidney cells (BHK21) with adenovirus type 12. Virology. 1969 Aug;38(4):587–606. doi: 10.1016/0042-6822(69)90179-2. [DOI] [PubMed] [Google Scholar]
  9. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Garner M. M., Revzin A. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res. 1981 Jul 10;9(13):3047–3060. doi: 10.1093/nar/9.13.3047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guggenheimer R. A., Nagata K., Lindenbaum J., Hurwitz J. Protein-primed replication of plasmids containing the terminus of the adenovirus genome. I. Characterization of an in vitro DNA replication system dependent on adenoviral DNA sequences. J Biol Chem. 1984 Jun 25;259(12):7807–7814. [PubMed] [Google Scholar]
  12. Harris M. P., Hay R. T. DNA sequences required for the initiation of adenovirus type 4 DNA replication in vitro. J Mol Biol. 1988 May 5;201(1):57–67. doi: 10.1016/0022-2836(88)90438-x. [DOI] [PubMed] [Google Scholar]
  13. Hatfield L., Hearing P. The NFIII/OCT-1 binding site stimulates adenovirus DNA replication in vivo and is functionally redundant with adjacent sequences. J Virol. 1993 Jul;67(7):3931–3939. doi: 10.1128/jvi.67.7.3931-3939.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hay R. T., Freeman A., Leith I., Monaghan A., Webster A. Molecular interactions during adenovirus DNA replication. Curr Top Microbiol Immunol. 1995;199(Pt 2):31–48. doi: 10.1007/978-3-642-79499-5_2. [DOI] [PubMed] [Google Scholar]
  15. Hay R. T., McDougall I. M. Viable viruses with deletions in the left inverted terminal repeat define the adenovirus origin of DNA replication. J Gen Virol. 1986 Feb;67(Pt 2):321–332. doi: 10.1099/0022-1317-67-2-321. [DOI] [PubMed] [Google Scholar]
  16. Hay R. T. Origin of adenovirus DNA replication. Role of the nuclear factor I binding site in vivo. J Mol Biol. 1985 Nov 5;186(1):129–136. doi: 10.1016/0022-2836(85)90263-3. [DOI] [PubMed] [Google Scholar]
  17. Klimkait T., Doerfler W. Adenovirus types 2 and 5 functions elicit replication and late expression of adenovirus type 12 DNA in hamster cells. J Virol. 1985 Aug;55(2):466–474. doi: 10.1128/jvi.55.2.466-474.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leegwater P. A., van Driel W., van der Vliet P. C. Recognition site of nuclear factor I, a sequence-specific DNA-binding protein from HeLa cells that stimulates adenovirus DNA replication. EMBO J. 1985 Jun;4(6):1515–1521. doi: 10.1002/j.1460-2075.1985.tb03811.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Leza M. A., Hearing P. Cellular transcription factor binds to adenovirus early region promoters and to a cyclic AMP response element. J Virol. 1988 Aug;62(8):3003–3013. doi: 10.1128/jvi.62.8.3003-3013.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lucher L. A., Khuntirat B., Zhao J., Angeletti P. C. Altered expression of adenovirus 12 DNA-binding protein but not DNA polymerase during abortive infection of hamster cells. Virology. 1992 Jul;189(1):187–195. doi: 10.1016/0042-6822(92)90694-k. [DOI] [PubMed] [Google Scholar]
  21. Miralles V. J., Cortes P., Stone N., Reinberg D. The adenovirus inverted terminal repeat functions as an enhancer in a cell-free system. J Biol Chem. 1989 Jun 25;264(18):10763–10772. [PubMed] [Google Scholar]
  22. Mul Y. M., Van der Vliet P. C. Nuclear factor I enhances adenovirus DNA replication by increasing the stability of a preinitiation complex. EMBO J. 1992 Feb;11(2):751–760. doi: 10.1002/j.1460-2075.1992.tb05108.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mul Y. M., Verrijzer C. P., van der Vliet P. C. Transcription factors NFI and NFIII/oct-1 function independently, employing different mechanisms to enhance adenovirus DNA replication. J Virol. 1990 Nov;64(11):5510–5518. doi: 10.1128/jvi.64.11.5510-5518.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nagata K., Guggenheimer R. A., Hurwitz J. Adenovirus DNA replication in vitro: synthesis of full-length DNA with purified proteins. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4266–4270. doi: 10.1073/pnas.80.14.4266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nagata K., Guggenheimer R. A., Hurwitz J. Specific binding of a cellular DNA replication protein to the origin of replication of adenovirus DNA. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6177–6181. doi: 10.1073/pnas.80.20.6177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Neill E. A., Fletcher C., Burrow C. R., Heintz N., Roeder R. G., Kelly T. J. Transcription factor OTF-1 is functionally identical to the DNA replication factor NF-III. Science. 1988 Sep 2;241(4870):1210–1213. doi: 10.1126/science.3413485. [DOI] [PubMed] [Google Scholar]
  27. O'Neill E. A., Kelly T. J. Purification and characterization of nuclear factor III (origin recognition protein C), a sequence-specific DNA binding protein required for efficient initiation of adenovirus DNA replication. J Biol Chem. 1988 Jan 15;263(2):931–937. [PubMed] [Google Scholar]
  28. Paonessa G., Gounari F., Frank R., Cortese R. Purification of a NF1-like DNA-binding protein from rat liver and cloning of the corresponding cDNA. EMBO J. 1988 Oct;7(10):3115–3123. doi: 10.1002/j.1460-2075.1988.tb03178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pruijn G. J., van Driel W., van der Vliet P. C. Nuclear factor III, a novel sequence-specific DNA-binding protein from HeLa cells stimulating adenovirus DNA replication. Nature. 1986 Aug 14;322(6080):656–659. doi: 10.1038/322656a0. [DOI] [PubMed] [Google Scholar]
  30. Pruijn G. J., van Miltenburg R. T., Claessens J. A., van der Vliet P. C. Interaction between the octamer-binding protein nuclear factor III and the adenovirus origin of DNA replication. J Virol. 1988 Sep;62(9):3092–3102. doi: 10.1128/jvi.62.9.3092-3102.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rosenfeld P. J., Kelly T. J. Purification of nuclear factor I by DNA recognition site affinity chromatography. J Biol Chem. 1986 Jan 25;261(3):1398–1408. [PubMed] [Google Scholar]
  32. Rosenfeld P. J., O'Neill E. A., Wides R. J., Kelly T. J. Sequence-specific interactions between cellular DNA-binding proteins and the adenovirus origin of DNA replication. Mol Cell Biol. 1987 Feb;7(2):875–886. doi: 10.1128/mcb.7.2.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Schiedner G., Schmitz B., Doerfler W. Late transcripts of adenovirus type 12 DNA are not translated in hamster cells expressing the E1 region of adenovirus type 5. J Virol. 1994 Sep;68(9):5476–5482. doi: 10.1128/jvi.68.9.5476-5482.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sprengel J., Schmitz B., Heuss-Neitzel D., Zock C., Doerfler W. Nucleotide sequence of human adenovirus type 12 DNA: comparative functional analysis. J Virol. 1994 Jan;68(1):379–389. doi: 10.1128/jvi.68.1.379-389.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stillman B. Initiation of eukaryotic DNA replication in vitro. Annu Rev Cell Biol. 1989;5:197–245. doi: 10.1146/annurev.cb.05.110189.001213. [DOI] [PubMed] [Google Scholar]
  37. Temperley S. M., Burrow C. R., Kelly T. J., Hay R. T. Identification of two distinct regions within the adenovirus minimal origin of replication that are required for adenovirus type 4 DNA replication in vitro. J Virol. 1991 Sep;65(9):5037–5044. doi: 10.1128/jvi.65.9.5037-5044.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Temperley S. M., Hay R. T. Replication of adenovirus type 4 DNA by a purified fraction from infected cells. Nucleic Acids Res. 1991 Jun 25;19(12):3243–3249. doi: 10.1093/nar/19.12.3243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tolun A., Aleström P., Pettersson U. Sequence of inverted terminal repetitions from different adenoviruses: demonstration of conserved sequences and homology between SA7 termini and SV40 DNA. Cell. 1979 Jul;17(3):705–713. doi: 10.1016/0092-8674(79)90277-0. [DOI] [PubMed] [Google Scholar]
  40. Van der Vliet P. C. Adenovirus DNA replication. Curr Top Microbiol Immunol. 1995;199(Pt 2):1–30. doi: 10.1007/978-3-642-79499-5_1. [DOI] [PubMed] [Google Scholar]
  41. Visser L., van Maarschalkerweerd M. W., Rozijn T. H., Wassenaar A. D., Reemst A. M., Sussenbach J. S. Viral DNA sequences in adenovirus-transformed cells. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):541–550. doi: 10.1101/sqb.1980.044.01.056. [DOI] [PubMed] [Google Scholar]
  42. Zock C., Doerfler W. A mitigator sequence in the downstream region of the major late promoter of adenovirus type 12 DNA. EMBO J. 1990 May;9(5):1615–1623. doi: 10.1002/j.1460-2075.1990.tb08281.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zock C., Iselt A., Doerfler W. A unique mitigator sequence determines the species specificity of the major late promoter in adenovirus type 12 DNA. J Virol. 1993 Feb;67(2):682–693. doi: 10.1128/jvi.67.2.682-693.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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