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. 1979 Feb;29(2):763–769. doi: 10.1128/jvi.29.2.763-769.1979

Transformation of human embryonic kidney cells by human papovarirus BK.

A F Purchio, G C Fareed
PMCID: PMC353208  PMID: 219257

Abstract

Infection of secondary human embryonic kidney (HEK) cells with human papovavirus BK (BKV) resulted in cellular lysis and degeneration within 7 days. After 30 days, multilayered colonies of transformed cells were found and subcultured for analyses. These BK-HEK cells uniformly expressed the BKV T-antigen but were only 1% V-antigen positive. They produced infectious virus and were resistant to superinfection by BKV. They reached a saturation density of 1.3 x 10(5) cells per cm2 in medium with 5% fetal calf serum, were able to grow in medium containing 2% serum, and did not form colonies in soft agar or tumors in nude mice. Nonintegrated, superhelical BKV DNA was detected in the noncloned cells as expected because they were persistently infected and contained RNA transcripts complementary to both early and late regions of the BKV genome. Analysis of T-antigen-positive clonal isolates of these BK-HEK cells by the Southern technique revealed an absence of free viral DNA and the presencce of integrated BKV DNA sequences corresponding to the early region of the BKV genome. These studies demonstrate the stable transformation of human cells by BKV. However, the transformed human cells which retain and express part of the BKV genome do not fully manifest the growth properties of other papovarirus-transformed cells.

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Selected References

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

  1. Birg F., Favaloro J., Kamen R. Analysis of polyoma virus nuclear RNA by mini-blot hybridization. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3138–3142. doi: 10.1073/pnas.74.8.3138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Botchan M., Topp W., Sambrook J. The arrangement of simian virus 40 sequences in the DNA of transformed cells. Cell. 1976 Oct;9(2):269–287. doi: 10.1016/0092-8674(76)90118-5. [DOI] [PubMed] [Google Scholar]
  3. Bouck N., Beales N., Shenk T., Berg P., di Mayorca G. New region of the simian virus 40 genome required for efficient viral transformation. Proc Natl Acad Sci U S A. 1978 May;75(5):2473–2477. doi: 10.1073/pnas.75.5.2473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradley M. K., Dougherty R. M. Transformation of African green monkey kidney cells with the RF strain of human papovavirus BKV. Virology. 1978 Mar;85(1):231–240. doi: 10.1016/0042-6822(78)90427-0. [DOI] [PubMed] [Google Scholar]
  5. Dhar R., Lai C. J., Khoury G. Nucleotide sequence of the DNA replication origin for human papovavirus BKV: sequence and structural homology with SV40. Cell. 1978 Feb;13(2):345–358. doi: 10.1016/0092-8674(78)90203-9. [DOI] [PubMed] [Google Scholar]
  6. Ferdinand F. J., Brown M., Khoury G. Characterization of early simian virus 40 transcriptional complexes: late transcription in the absence of detectable DNA replication. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5443–5447. doi: 10.1073/pnas.74.12.5443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fiori M., Di Mayorca G. Occurrence of BK virus DNA in DNA obtained from certain human tumors. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4662–4666. doi: 10.1073/pnas.73.12.4662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gardner S. D., Field A. M., Coleman D. V., Hulme B. New human papovavirus (B.K.) isolated from urine after renal transplantation. Lancet. 1971 Jun 19;1(7712):1253–1257. doi: 10.1016/s0140-6736(71)91776-4. [DOI] [PubMed] [Google Scholar]
  9. Gardner S. D. Prevalence in England of antibody to human polyomavirus (B.k.). Br Med J. 1973 Jan 13;1(5845):77–78. doi: 10.1136/bmj.1.5845.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Howley P. M., Khoury G., Byrne J. C., Takemoto K. K., Martin M. A. Physical map of the BK virus genome. J Virol. 1975 Oct;16(4):959–973. doi: 10.1128/jvi.16.4.959-973.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Howley P. M., Mullarkey M. F., Takemoto K. K., Martin M. A. Characterization of human papovavirus BK DNA. J Virol. 1975 Jan;15(1):173–181. doi: 10.1128/jvi.15.1.173-181.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lodish H. F., Jacobson A., Firtel R., Alton T., Tuchman J. Synthesis of messenger RNA and chromosome structure in the cellular slime mold. Proc Natl Acad Sci U S A. 1974 Dec;71(12):5103–5108. doi: 10.1073/pnas.71.12.5103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Major E. O., Di Mayorca G. Malignant transformation of BHK21 clone 13 cells by BK virus--a human papovavirus. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3210–3212. doi: 10.1073/pnas.70.11.3210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maniatis T., Kee S. G., Efstratiadis A., Kafatos F. C. Amplification and characterization of a beta-globin gene synthesized in vitro. Cell. 1976 Jun;8(2):163–182. doi: 10.1016/0092-8674(76)90001-5. [DOI] [PubMed] [Google Scholar]
  17. Martinis J., Croce C. M. Somatic cell hybrids producing antibodies specific for the tumor antigen of simian virus 40. Proc Natl Acad Sci U S A. 1978 May;75(5):2320–2323. doi: 10.1073/pnas.75.5.2320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mullarkey M. F., Hruska J. F., Takemoto K. K. Comparison of two human papovaviruses with simian virus 40 by structural protein and antigenic analysis. J Virol. 1974 May;13(5):1014–1019. doi: 10.1128/jvi.13.5.1014-1019.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Newell N., Lai C. J., Khoury G., Kelly T. J., Jr Electron microscope study of the base sequence homology between simian virus 40 and human papovavirus BK. J Virol. 1978 Jan;25(1):193–201. doi: 10.1128/jvi.25.1.193-201.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Osborn J. E., Robertson S. M., Padgett B. L., Walker D. L., Weisblum B. Comparison of JC and BK human papovaviruses with simian virus 40: DNA homology studies. J Virol. 1976 Aug;19(2):675–684. doi: 10.1128/jvi.19.2.675-684.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Portolani M., Barbanti-Brodano G., Placa M. L. Malignant transformation of hamster kidney cells by BK virus. J Virol. 1975 Feb;15(2):420–422. doi: 10.1128/jvi.15.2.420-422.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Simmons D. T., Takemoto K. K., Martin M. A. Relationship between the methionine tryptic peptides of simian virus 40 and BK virus tumor antigens. J Virol. 1977 Oct;24(1):319–325. doi: 10.1128/jvi.24.1.319-325.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sleigh M. J., Topp W. C., Hanich R., Sambrook J. F. Mutants of SV40 with an altered small t protein are reduced in their ability to transform cells. Cell. 1978 May;14(1):79–88. doi: 10.1016/0092-8674(78)90303-3. [DOI] [PubMed] [Google Scholar]
  24. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  25. Takemoto K. K., Martin M. A. Transformation of hamster kidney cells by BK papovavirus DNA. J Virol. 1975 Jan;17(1):247–253. doi: 10.1128/jvi.17.1.247-253.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Takemoto K. K., Mullarkey M. F. Human papovavirus, BK strain: biological studies including antigenic relationship to simian virus 40. J Virol. 1973 Sep;12(3):625–631. doi: 10.1128/jvi.12.3.625-631.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Upcroft P., Skolnik H., Upcroft J. A., Solomon D., Khoury G., Hamer D. H., Fareed G. C. Transduction of a bacterial gene into mammalian cells. Proc Natl Acad Sci U S A. 1978 May;75(5):2117–2121. doi: 10.1073/pnas.75.5.2117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wilson G. A., Young F. E. Isolation of a sequence-specific endonuclease (BamI) from Bacillus amyloliquefaciens H. J Mol Biol. 1975 Sep 5;97(1):123–125. doi: 10.1016/s0022-2836(75)80028-3. [DOI] [PubMed] [Google Scholar]
  29. Wold W. S., Mackey J. K., Brackmann K. H., Takemori N., Rigden P., Green M. Analysis of human tumors and human malignant cell lines for BK virus-specific DNA sequences. Proc Natl Acad Sci U S A. 1978 Jan;75(1):454–458. doi: 10.1073/pnas.75.1.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yang R. C., Wu R. BK virus DNA: cleavage map and sequence analysis. Proc Natl Acad Sci U S A. 1978 May;75(5):2150–2154. doi: 10.1073/pnas.75.5.2150. [DOI] [PMC free article] [PubMed] [Google Scholar]

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