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. 1980 Mar;33(3):1225–1228. doi: 10.1128/jvi.33.3.1225-1228.1980

Integration Pattern of Human JC Virus Sequences in Two Clones of a Cell Line Established from a JC Virus-Induced Hamster Brain Tumor

William S M Wold 1, Maurice Green 1, Jesse K Mackey 1, Jonathan D Martin 2, Billie L Padgett 2, Duard L Walker 2
PMCID: PMC288659  PMID: 6245274

Abstract

The physical state of the JC virus (JCV) genome was studied in two clonal cell lines (clones 2 and 7) derived from a tissue culture cell line (HJC-15) established from a hamster brain tumor induced by JCV. Saturation-hybridization and reassociation kinetic analyses, using in vitro 32P-labeled JCV DNA, indicated that clone 7 and 2 cells contain 9 to 10 and 4 to 5 copies per cell, respectively, of all or most of the viral genome. Both cell DNAs were analyzed by using the Southern blotting procedure with three restriction endonucleases: XhoI, which does not cleave JCV DNA; EcoRI, which cleaves once; and HindIII, which cleaves three times. With each DNA, a variety of JCV-specific DNA fragments were detected. The following conclusions are possible: (i) JCV DNA is integrated into cell DNA in both clonal lines; (ii) both clonal lines contain multiple copies of the viral genome integrated in a tandem head-to-tail orientation; (iii) neither clonal line contains detectable free-form I, II, or III JCV DNA; (iv) each clonal line contains multiple independent sites of JCV DNA integration; and (v) most or all of the sites of integration on the cellular or the viral genome, or both, are different in clone 7 DNA than in clone 2 DNA. Thus, although both clone 7 and clone 2 cells were established from the HJC-15 tumor cell line, they differ in the copy number and integration pattern of JCV DNA.

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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., Dulbecco R., Fried M., Kamen R. State and organization of polyoma virus DNA sequences in transformed rat cell lines. J Virol. 1979 Feb;29(2):633–648. doi: 10.1128/jvi.29.2.633-648.1979. [DOI] [PMC free article] [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. 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]
  4. Campo M. S., Cameron I. R., Rogers M. E. Tandem integration of complete and defective SV40 genomes in mouse-human somatic cell hybrids. Cell. 1978 Dec;15(4):1411–1426. doi: 10.1016/0092-8674(78)90065-x. [DOI] [PubMed] [Google Scholar]
  5. Graham D. E. The isolation of high molecular weight DNA from whole organisms or large tissue masses. Anal Biochem. 1978 Apr;85(2):609–613. doi: 10.1016/0003-2697(78)90262-2. [DOI] [PubMed] [Google Scholar]
  6. Howley P. M., Khoury G., Takemoto K. K., Martin M. A. Polynucleotide sequences common to the genomes of simian virus 40 and the human papovaviruses JC and BK. Virology. 1976 Aug;73(1):303–307. doi: 10.1016/0042-6822(76)90084-2. [DOI] [PubMed] [Google Scholar]
  7. Ketner G., Kelly T. J., Jr Integrated simian virus 40 sequences in transformed cell DNA: analysis using restriction endonucleases. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1102–1106. doi: 10.1073/pnas.73.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mackey J. K., Brackmann K. H., Green M. R., Green M. Preparation and characterization of highly radioactive in vitro labeled adenovirus DNA and DNA restriction fragments. Biochemistry. 1977 Oct 4;16(20):4478–4483. doi: 10.1021/bi00639a023. [DOI] [PubMed] [Google Scholar]
  9. Martin J. D., Frisque R. J., Padgett B. L., Walker D. L. Restriction endonuclease cleavage map of the DNA of JC virus. J Virol. 1979 Mar;29(3):846–855. doi: 10.1128/jvi.29.3.846-855.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Osborn J. E., Robertson S. M., Padgett B. L., ZuRhein G. M., Walker D. L., Weisblum B. Comparison of JC and BK human papovaviruses with simian virus 40: restriction endonuclease digestion and gel electrophoresis of resultant fragments. J Virol. 1974 Mar;13(3):614–622. doi: 10.1128/jvi.13.3.614-622.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Padgett B. L., Walker D. L. New human papovaviruses. Prog Med Virol. 1976;22:1–35. [PubMed] [Google Scholar]
  13. Padgett B. L., Walker D. L., ZuRhein G. M., Eckroade R. J., Dessel B. H. Cultivation of papova-like virus from human brain with progressive multifocal leucoencephalopathy. Lancet. 1971 Jun 19;1(7712):1257–1260. doi: 10.1016/s0140-6736(71)91777-6. [DOI] [PubMed] [Google Scholar]
  14. Rigby P. W., Berg P. Does simian virus 40 DNA integrate into cellular DNA during productive infection? J Virol. 1978 Nov;28(2):475–489. doi: 10.1128/jvi.28.2.475-489.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Steinberg B., Pollack R., Topp W., Botchan M. Isolation and characterization of T antigen-negative revertants from a line of transformed rat cells containing one copy of the SV40 genome. Cell. 1978 Jan;13(1):19–32. doi: 10.1016/0092-8674(78)90134-4. [DOI] [PubMed] [Google Scholar]
  17. Takemoto K. K., Howley P. M., Miyamura T. JC human papovavirus replication in human amnion cells. J Virol. 1979 Apr;30(1):384–389. doi: 10.1128/jvi.30.1.384-389.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Walker D. L., Padgett B. L., ZuRhein G. M., Albert A. E., Marsh R. F. Human papovavirus (JC): induction of brain tumors in hamsters. Science. 1973 Aug 17;181(4100):674–676. doi: 10.1126/science.181.4100.674. [DOI] [PubMed] [Google Scholar]

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