Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. May 1989; 63(5): 2180–2190.
PMCID: PMC250635

JC virus-simian virus 40 genomes containing heterologous regulatory signals and chimeric early regions: identification of regions restricting transformation by JC virus.


The papovavirus JC virus (JCV) is highly oncogenic in experimental animals but, unlike simian virus 40 (SV40), is severely restricted in its ability to transform cells in culture. We exploited the close genetic relatedness of these two viruses to delimit region(s) of the T protein which can restrict transforming activity. Novel chimeric genomes were produced by exchanging various segments of the JCV and SV40 T-protein-coding regions. These DNA constructs specified early proteins with in-frame substitutions of analogous amino acid sequences. A second set of genomes was prepared which, in addition to chimeric early proteins, contained substituted regulatory regions. The transformation efficiencies of these chimeric genomes were intermediate between those of SV40 and JCV, with the source of T protein exerting a greater effect than that of the regulatory region. The ability of certain constructs to induce efficient transformation required the presence of an SV40 regulatory region or specific sequences within the SV40 early coding region. Cloned cell lines prepared from representative transformants were characterized; the ability to form colonies in soft agarose was investigated, and the presence of viral T and cellular p53 proteins was determined. The various T proteins differed in amount, stability, and the ability to form stable complexes with p53.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anton IA, Lane DP. Non-structural protein 1 of parvoviruses: homology to purine nucleotide using proteins and early proteins of papovaviruses. Nucleic Acids Res. 1986 Oct 10;14(19):7813–7813. [PMC free article] [PubMed]
  • Arthur AK, Höss A, Fanning E. Expression of simian virus 40 T antigen in Escherichia coli: localization of T-antigen origin DNA-binding domain to within 129 amino acids. J Virol. 1988 Jun;62(6):1999–2006. [PMC free article] [PubMed]
  • Asselin C, Bastin M. Sequences from polyomavirus and simian virus 40 large T genes capable of immortalizing primary rat embryo fibroblasts. J Virol. 1985 Dec;56(3):958–968. [PMC free article] [PubMed]
  • Beth E, Cikes M, Schloen L, di Mayorca G, Giraldo G. Interspecies-, species- and type-specific T antigenic determinants of human papovaviruses (JC and BK) and of Simian virus 40. Int J Cancer. 1977 Oct 15;20(4):551–559. [PubMed]
  • Bikel I, Montano X, Agha ME, Brown M, McCormack M, Boltax J, Livingston DM. SV40 small t antigen enhances the transformation activity of limiting concentrations of SV40 large T antigen. Cell. 1987 Jan 30;48(2):321–330. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Bollag B, Chuke WF, Frisque RJ. Hybrid genomes of the polyomaviruses JC virus, BK virus, and simian virus 40: identification of sequences important for efficient transformation. J Virol. 1989 Feb;63(2):863–872. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Bradley MK, Hudson J, Villanueva MS, Livingston DM. Specific in vitro adenylylation of the simian virus 40 large tumor antigen. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6574–6578. [PMC free article] [PubMed]
  • Bradley MK, Smith TF, Lathrop RH, Livingston DM, Webster TA. Consensus topography in the ATP binding site of the simian virus 40 and polyomavirus large tumor antigens. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4026–4030. [PMC free article] [PubMed]
  • Chang LS, Pan S, Pater MM, Di Mayorca G. Differential requirement for SV40 early genes in immortalization and transformation of primary rat and human embryonic cells. Virology. 1985 Oct 30;146(2):246–261. [PubMed]
  • Cherington V, Brown M, Paucha E, St Louis J, Spiegelman BM, Roberts TM. Separation of simian virus 40 large-T-antigen-transforming and origin-binding functions from the ability to block differentiation. Mol Cell Biol. 1988 Mar;8(3):1380–1384. [PMC free article] [PubMed]
  • Chesters PM, Heritage J, McCance DJ. Persistence of DNA sequences of BK virus and JC virus in normal human tissues and in diseased tissues. J Infect Dis. 1983 Apr;147(4):676–684. [PubMed]
  • Chuke WF, Walker DL, Peitzman LB, Frisque RJ. Construction and characterization of hybrid polyomavirus genomes. J Virol. 1986 Dec;60(3):960–971. [PMC free article] [PubMed]
  • Clayton CE, Murphy D, Lovett M, Rigby PW. A fragment of the SV40 large T-antigen gene transforms. Nature. 1982 Sep 2;299(5878):59–61. [PubMed]
  • Clertant P, Gaudray P, May E, Cuzin F. The nucleotide binding site detected by affinity labeling in the large T proteins of polyoma and SV40 viruses is distinct from their ATPase catalytic site. J Biol Chem. 1984 Dec 25;259(24):15196–15203. [PubMed]
  • Colby WW, Shenk T. Fragments of the simian virus 40 transforming gene facilitate transformation of rat embryo cells. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5189–5193. [PMC free article] [PubMed]
  • Coleman DV, Wolfendale MR, Daniel RA, Dhanjal NK, Gardner SD, Gibson PE, Field AM. A prospective study of human polyomavirus infection in pregnancy. J Infect Dis. 1980 Jul;142(1):1–8. [PubMed]
  • Coussens PM, Cooper JA, Hunter T, Shalloway D. Restriction of the in vitro and in vivo tyrosine protein kinase activities of pp60c-src relative to pp60v-src. Mol Cell Biol. 1985 Oct;5(10):2753–2763. [PMC free article] [PubMed]
  • DeCaprio JA, Ludlow JW, Figge J, Shew JY, Huang CM, Lee WH, Marsilio E, Paucha E, Livingston DM. SV40 large tumor antigen forms a specific complex with the product of the retinoblastoma susceptibility gene. Cell. 1988 Jul 15;54(2):275–283. [PubMed]
  • Figge J, Webster T, Smith TF, Paucha E. Prediction of similar transforming regions in simian virus 40 large T, adenovirus E1A, and myc oncoproteins. J Virol. 1988 May;62(5):1814–1818. [PMC free article] [PubMed]
  • Frisque RJ. Regulatory sequences and virus-cell interactions of JC virus. Prog Clin Biol Res. 1983;105:41–59. [PubMed]
  • Frisque RJ, Bream GL, Cannella MT. Human polyomavirus JC virus genome. J Virol. 1984 Aug;51(2):458–469. [PMC free article] [PubMed]
  • Frisque RJ, Rifkin DB, Topp WC. Requirement for the large T and small T proteins of SV40 in the maintenance of the transformed state. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):325–331. [PubMed]
  • Gurney EG, Harrison RO, Fenno J. Monoclonal antibodies against simian virus 40 T antigens: evidence for distinct sublcasses of large T antigen and for similarities among nonviral T antigens. J Virol. 1980 Jun;34(3):752–763. [PMC free article] [PubMed]
  • Hara K, Oya Y, Yogo Y. Enhancement of the transforming capacity of BK virus by partial deletion of the 68-base-pair tandem repeats. J Virol. 1985 Sep;55(3):867–869. [PMC free article] [PubMed]
  • Hogan TF, Borden EC, McBain JA, Padgett BL, Walker DL. Human polyomavirus infections with JC virus and BK virus in renal transplant patients. Ann Intern Med. 1980 Mar;92(3):373–378. [PubMed]
  • Houff SA, Major EO, Katz DA, Kufta CV, Sever JL, Pittaluga S, Roberts JR, Gitt J, Saini N, Lux W. Involvement of JC virus-infected mononuclear cells from the bone marrow and spleen in the pathogenesis of progressive multifocal leukoencephalopathy. N Engl J Med. 1988 Feb 4;318(5):301–305. [PubMed]
  • Kalderon D, Smith AE. In vitro mutagenesis of a putative DNA binding domain of SV40 large-T. Virology. 1984 Nov;139(1):109–137. [PubMed]
  • Kenney S, Natarajan V, Strike D, Khoury G, Salzman NP. JC virus enhancer-promoter active in human brain cells. Science. 1984 Dec 14;226(4680):1337–1339. [PubMed]
  • Kriegler M, Botchan M. Enhanced transformation by a simian virus 40 recombinant virus containing a Harvey murine sarcoma virus long terminal repeat. Mol Cell Biol. 1983 Mar;3(3):325–339. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Linzer DI, Maltzman W, Levine AJ. The SV40 A gene product is required for the production of a 54,000 MW cellular tumor antigen. Virology. 1979 Oct 30;98(2):308–318. [PubMed]
  • Livingston DM, Bradley MK. The simian virus 40 large T antigen. A lot packed into a little. Mol Biol Med. 1987 Apr;4(2):63–80. [PubMed]
  • London WT, Houff SA, Madden DL, Fuccillo DA, Gravell M, Wallen WC, Palmer AE, Sever JL, Padgett BL, Walker DL, et al. Brain tumors in owl monkeys inoculated with a human polyomavirus (JC virus). Science. 1978 Sep 29;201(4362):1246–1249. [PubMed]
  • Marusyk R, Sergeant A. A simple method for dialysis of small-volume samples. Anal Biochem. 1980 Jul 1;105(2):403–404. [PubMed]
  • Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. [PubMed]
  • Mercer WE, Nelson D, DeLeo AB, Old LJ, Baserga R. Microinjection of monoclonal antibody to protein p53 inhibits serum-induced DNA synthesis in 3T3 cells. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6309–6312. [PMC free article] [PubMed]
  • Michalovitz D, Eliyahu D, Oren M. Overproduction of protein p53 contributes to simian virus 40-mediated transformation. Mol Cell Biol. 1986 Oct;6(10):3531–3536. [PMC free article] [PubMed]
  • Miyamura T, Jikuya H, Soeda E, Yoshiike K. Genomic structure of human polyoma virus JC: nucleotide sequence of the region containing replication origin and small-T-antigen gene. J Virol. 1983 Jan;45(1):73–79. [PMC free article] [PubMed]
  • Mole SE, Gannon JV, Ford MJ, Lane DP. Structure and function of SV40 large-T antigen. Philos Trans R Soc Lond B Biol Sci. 1987 Dec 15;317(1187):455–469. [PubMed]
  • Moran E. A region of SV40 large T antigen can substitute for a transforming domain of the adenovirus E1A products. Nature. 1988 Jul 14;334(6178):168–170. [PubMed]
  • Oren M, Maltzman W, Levine AJ. Post-translational regulation of the 54K cellular tumor antigen in normal and transformed cells. Mol Cell Biol. 1981 Feb;1(2):101–110. [PMC free article] [PubMed]
  • Ohsumi S, Motoi M, Ogawa K. Induction of undifferentiated tumors by JC virus in the cerebrum of rats. Acta Pathol Jpn. 1986 Jun;36(6):815–825. [PubMed]
  • Padgett BL, Walker DL. Prevalence of antibodies in human sera against JC virus, an isolate from a case of progressive multifocal leukoencephalopathy. J Infect Dis. 1973 Apr;127(4):467–470. [PubMed]
  • Padgett BL, Walker DL, ZuRhein GM, Varakis JN. Differential neurooncogenicity of strains of JC virus, a human polyoma virus, in newborn Syrian hamsters. Cancer Res. 1977 Mar;37(3):718–720. [PubMed]
  • Pan S, Sompayrac LM, Knowles BB, Danna KJ. A fragment of the simian virus 40 early genome can induce tumors in nude mice. J Virol. 1985 Mar;53(3):988–989. [PMC free article] [PubMed]
  • Paucha E, Kalderon D, Harvey RW, Smith AE. Simian virus 40 origin DNA-binding domain on large T antigen. J Virol. 1986 Jan;57(1):50–64. [PMC free article] [PubMed]
  • Reich NC, Levine AJ. Growth regulation of a cellular tumour antigen, p53, in nontransformed cells. Nature. 1984 Mar 8;308(5955):199–201. [PubMed]
  • Rieth KG, Di Chiro G, London WT, Sever JL, Houff SA, Kornblith PL, McKeever PE, Buonomo C, Padgett BL, Walker DL. Experimental glioma in primates: a computed tomography model. J Comput Assist Tomogr. 1980 Jun;4(3):285–290. [PubMed]
  • Shenk TE, Carbon J, Berg P. Construction and analysis of viable deletion mutants of simian virus 40. J Virol. 1976 May;18(2):664–671. [PMC free article] [PubMed]
  • Sleigh MJ, Topp WC, Hanich R, Sambrook JF. Mutants of SV40 with an altered small t protein are reduced in their ability to transform cells. Cell. 1978 May;14(1):79–88. [PubMed]
  • Small MB, Gluzman Y, Ozer HL. Enhanced transformation of human fibroblasts by origin-defective simian virus 40. Nature. 1982 Apr 15;296(5858):671–672. [PubMed]
  • Sompayrac L, Danna KJ. Less than 40% of the simian virus 40 large T-antigen-coding sequence is required for transformation. Mol Cell Biol. 1984 Aug;4(8):1661–1663. [PMC free article] [PubMed]
  • Sompayrac L, Danna KJ. The simian virus 40 sequences between 0.169 and 0.423 map units are not essential to immortalize early-passage rat embryo cells. Mol Cell Biol. 1985 May;5(5):1191–1194. [PMC free article] [PubMed]
  • Sompayrac L, Danna KJ. A new SV40 mutant that encodes a small fragment of T antigen transforms established rat and mouse cells. Virology. 1988 Apr;163(2):391–396. [PubMed]
  • Soprano KJ, Galanti N, Jonak GJ, McKercher S, Pipas JM, Peden KW, Baserga R. Mutational analysis of simian virus 40 T antigen: stimulation of cellular DNA synthesis and activation of rRNA genes by mutants with deletions in the T-antigen gene. Mol Cell Biol. 1983 Feb;3(2):214–219. [PMC free article] [PubMed]
  • Tack LC, Wright JH, Gurney EG. Characterization of simian virus 40 large T antigen by using different monoclonal antibodies: T-p53 complexes are preferentially ATPase active and adenylylated. J Virol. 1988 Mar;62(3):1028–1037. [PMC free article] [PubMed]
  • Tordaro GJ, Green H. An assay for cellular transformation by SV40. Virology. 1964 May;23(1):117–119. [PubMed]
  • Topp WC. Normal rat cell lines deficient in nuclear thymidine kinase. Virology. 1981 Aug;113(1):408–411. [PubMed]
  • Tornow J, Polvino-Bodnar M, Santangelo G, Cole CN. Two separable functional domains of simian virus 40 large T antigen: carboxyl-terminal region of simian virus 40 large T antigen is required for efficient capsid protein synthesis. J Virol. 1985 Feb;53(2):415–424. [PMC free article] [PubMed]
  • Walker DL, Padgett BL, ZuRhein GM, Albert AE, Marsh RF. Human papovavirus (JC): induction of brain tumors in hamsters. Science. 1973 Aug 17;181(4100):674–676. [PubMed]
  • Watanabe S, Soeda E, Uchida S, Yoshiike K. DNA rearrangement affecting expression of the BK virus transforming gene. J Virol. 1984 Jul;51(1):1–6. [PMC free article] [PubMed]
  • Watanabe S, Yoshiike K. Decreasing the number of 68-base-pair tandem repeats in the BK virus transcriptional control region reduces plaque size and enhances transforming capacity. J Virol. 1985 Sep;55(3):823–825. [PMC free article] [PubMed]
  • Wigler M, Pellicer A, Silverstein S, Axel R. Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978 Jul;14(3):725–731. [PubMed]

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


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...