Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1997 Oct;71(10):7305–7311. doi: 10.1128/jvi.71.10.7305-7311.1997

Human foamy virus reverse transcription that occurs late in the viral replication cycle.

A Moebes 1, J Enssle 1, P D Bieniasz 1, M Heinkelein 1, D Lindemann 1, M Bock 1, M O McClure 1, A Rethwilm 1
PMCID: PMC192074  PMID: 9311807

Abstract

Foamy viruses (FVs) are retroid viruses which use a replication strategy unlike those of other retroviruses and hepadnaviruses (S. F. Yu, D. N. Baldwin, S. R. Gwynn, S. Yendapilli, and M. L. Linial, Science 271:1579-1582, 1996). One of the striking differences between FVs and retroviruses is the presence of large amounts of linear genome-length DNA in FV-infected cells and in virions. We report here that large quantities of genome-length linear FV DNA accumulate in cells infected with FV, as determined by Southern blotting. To determine whether these unintegrated virus DNAs result solely from superinfection, we analyzed the occurrence of virus cDNA of the so-called human FV isolate (HFV) in cells transfected with a virus mutant deficient in the envelope gene and in cells which are resistant to superinfection due to stable expression of the envelope protein. We show that the synthesis of viral cDNA is independent of superinfection and that HFV synthesizes cDNA intracellularly as a late event in the replication cycle. To further confirm this finding, we performed inhibition studies with the reverse transcriptase inhibitor zidovudine (AZT). While AZT had no effect or only a minor effect on virus titers when added to cells prior to virus infection, viral titers were reduced by 3 or 4 orders of magnitude when the virus was produced from cells in the presence of AZT. Our results are most compatible with the hypothesis that the functional nucleic acid of the extracellular HFV consists of largely double-stranded linear DNA.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Baunach G., Maurer B., Hahn H., Kranz M., Rethwilm A. Functional analysis of human foamy virus accessory reading frames. J Virol. 1993 Sep;67(9):5411–5418. doi: 10.1128/jvi.67.9.5411-5418.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benkirane M., Corbeau P., Housset V., Devaux C. An antibody that binds the immunoglobulin CDR3-like region of the CD4 molecule inhibits provirus transcription in HIV-infected T cells. EMBO J. 1993 Dec 15;12(13):4909–4921. doi: 10.1002/j.1460-2075.1993.tb06185.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bieniasz P. D., Rethwilm A., Pitman R., Daniel M. D., Chrystie I., McClure M. O. A comparative study of higher primate foamy viruses, including a new virus from a gorilla. Virology. 1995 Feb 20;207(1):217–228. doi: 10.1006/viro.1995.1068. [DOI] [PubMed] [Google Scholar]
  4. Chen I. S., Temin H. M. Establishment of infection by spleen necrosis virus: inhibition in stationary cells and the role of secondary infection. J Virol. 1982 Jan;41(1):183–191. doi: 10.1128/jvi.41.1.183-191.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Enssle J., Jordan I., Mauer B., Rethwilm A. Foamy virus reverse transcriptase is expressed independently from the Gag protein. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4137–4141. doi: 10.1073/pnas.93.9.4137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Furman P. A., Fyfe J. A., St Clair M. H., Weinhold K., Rideout J. L., Freeman G. A., Lehrman S. N., Bolognesi D. P., Broder S., Mitsuya H. Phosphorylation of 3'-azido-3'-deoxythymidine and selective interaction of the 5'-triphosphate with human immunodeficiency virus reverse transcriptase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8333–8337. doi: 10.1073/pnas.83.21.8333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goepfert P. A., Shaw K. L., Ritter G. D., Jr, Mulligan M. J. A sorting motif localizes the foamy virus glycoprotein to the endoplasmic reticulum. J Virol. 1997 Jan;71(1):778–784. doi: 10.1128/jvi.71.1.778-784.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goepfert P. A., Wang G., Mulligan M. J. Identification of an ER retrieval signal in a retroviral glycoprotein. Cell. 1995 Aug 25;82(4):543–544. doi: 10.1016/0092-8674(95)90026-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  10. Granowitz C., Colicelli J., Goff S. P. Analysis of mutations in the envelope gene of Moloney murine leukemia virus: separation of infectivity from superinfection resistance. Virology. 1991 Aug;183(2):545–554. doi: 10.1016/0042-6822(91)90983-i. [DOI] [PubMed] [Google Scholar]
  11. Harbers K., Schnieke A., Stuhlmann H., Jähner D., Jaenisch R. DNA methylation and gene expression: endogenous retroviral genome becomes infectious after molecular cloning. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7609–7613. doi: 10.1073/pnas.78.12.7609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jordan I., Enssle J., Güttler E., Mauer B., Rethwilm A. Expression of human foamy virus reverse transcriptase involves a spliced pol mRNA. Virology. 1996 Oct 1;224(1):314–319. doi: 10.1006/viro.1996.0534. [DOI] [PubMed] [Google Scholar]
  13. Kim S. Y., Byrn R., Groopman J., Baltimore D. Temporal aspects of DNA and RNA synthesis during human immunodeficiency virus infection: evidence for differential gene expression. J Virol. 1989 Sep;63(9):3708–3713. doi: 10.1128/jvi.63.9.3708-3713.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kupiec J. J., Tobaly-Tapiero J., Canivet M., Santillana-Hayat M., Flügel R. M., Périès J., Emanoil-Ravier R. Evidence for a gapped linear duplex DNA intermediate in the replicative cycle of human and simian spumaviruses. Nucleic Acids Res. 1988 Oct 25;16(20):9557–9565. doi: 10.1093/nar/16.20.9557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kögel D., Aboud M., Flügel R. M. Mutational analysis of the reverse transcriptase and ribonuclease H domains of the human foamy virus. Nucleic Acids Res. 1995 Jul 25;23(14):2621–2625. doi: 10.1093/nar/23.14.2621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Levinson W. E., Varmus H. E., Garapin A. C., Bishop J. M. DNA of Rous sarcoma virus: its nature and significance. Science. 1972 Jan 7;175(4017):76–78. doi: 10.1126/science.175.4017.76. [DOI] [PubMed] [Google Scholar]
  17. Lewis P., Hensel M., Emerman M. Human immunodeficiency virus infection of cells arrested in the cell cycle. EMBO J. 1992 Aug;11(8):3053–3058. doi: 10.1002/j.1460-2075.1992.tb05376.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lindemann D., Bock M., Schweizer M., Rethwilm A. Efficient pseudotyping of murine leukemia virus particles with chimeric human foamy virus envelope proteins. J Virol. 1997 Jun;71(6):4815–4820. doi: 10.1128/jvi.71.6.4815-4820.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lori F., di Marzo Veronese F., de Vico A. L., Lusso P., Reitz M. S., Jr, Gallo R. C. Viral DNA carried by human immunodeficiency virus type 1 virions. J Virol. 1992 Aug;66(8):5067–5074. doi: 10.1128/jvi.66.8.5067-5074.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mergia A., Luciw P. A. Replication and regulation of primate foamy viruses. Virology. 1991 Oct;184(2):475–482. doi: 10.1016/0042-6822(91)90417-a. [DOI] [PubMed] [Google Scholar]
  21. Mitsuya H., Broder S. Strategies for antiviral therapy in AIDS. 1987 Feb 26-Mar 4Nature. 325(6107):773–778. doi: 10.1038/325773a0. [DOI] [PubMed] [Google Scholar]
  22. Morgenstern J. P., Land H. Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res. 1990 Jun 25;18(12):3587–3596. doi: 10.1093/nar/18.12.3587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mullins J. I., Chen C. S., Hoover E. A. Disease-specific and tissue-specific production of unintegrated feline leukaemia virus variant DNA in feline AIDS. Nature. 1986 Jan 23;319(6051):333–336. doi: 10.1038/319333a0. [DOI] [PubMed] [Google Scholar]
  24. Nassal M., Schaller H. Hepatitis B virus replication. Trends Microbiol. 1993 Sep;1(6):221–228. doi: 10.1016/0966-842x(93)90136-f. [DOI] [PubMed] [Google Scholar]
  25. Rethwilm A. Unexpected replication pathways of foamy viruses. J Acquir Immune Defic Syndr Hum Retrovirol. 1996;13 (Suppl 1):S248–S253. doi: 10.1097/00042560-199600001-00037. [DOI] [PubMed] [Google Scholar]
  26. Saïb A., Puvion-Dutilleul F., Schmid M., Périès J., de Thé H. Nuclear targeting of incoming human foamy virus Gag proteins involves a centriolar step. J Virol. 1997 Feb;71(2):1155–1161. doi: 10.1128/jvi.71.2.1155-1161.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schliephake A. W., Rethwilm A. Nuclear localization of foamy virus Gag precursor protein. J Virol. 1994 Aug;68(8):4946–4954. doi: 10.1128/jvi.68.8.4946-4954.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schmidt M., Herchenröder O., Heeney J., Rethwilm A. Long terminal repeat U3 length polymorphism of human foamy virus. Virology. 1997 Apr 14;230(2):167–178. doi: 10.1006/viro.1997.8463. [DOI] [PubMed] [Google Scholar]
  29. Schmidt M., Rethwilm A. Replicating foamy virus-based vectors directing high level expression of foreign genes. Virology. 1995 Jun 20;210(1):167–178. doi: 10.1006/viro.1995.1328. [DOI] [PubMed] [Google Scholar]
  30. Schweizer M., Renne R., Neumann-Haefelin D. Structural analysis of proviral DNA in simian foamy virus (LK-3)-infected cells. Arch Virol. 1989;109(1-2):103–114. doi: 10.1007/BF01310521. [DOI] [PubMed] [Google Scholar]
  31. Soneoka Y., Cannon P. M., Ramsdale E. E., Griffiths J. C., Romano G., Kingsman S. M., Kingsman A. J. A transient three-plasmid expression system for the production of high titer retroviral vectors. Nucleic Acids Res. 1995 Feb 25;23(4):628–633. doi: 10.1093/nar/23.4.628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Temin H. M. Mechanisms of cell killing/cytopathic effects by nonhuman retroviruses. Rev Infect Dis. 1988 Mar-Apr;10(2):399–405. doi: 10.1093/clinids/10.2.399. [DOI] [PubMed] [Google Scholar]
  33. Trono D. Partial reverse transcripts in virions from human immunodeficiency and murine leukemia viruses. J Virol. 1992 Aug;66(8):4893–4900. doi: 10.1128/jvi.66.8.4893-4900.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Varmus H. E. Form and function of retroviral proviruses. Science. 1982 May 21;216(4548):812–820. doi: 10.1126/science.6177038. [DOI] [PubMed] [Google Scholar]
  35. Weiss R. A. Reverse transcription. Foamy viruses bubble on. Nature. 1996 Mar 21;380(6571):201–201. doi: 10.1038/380201a0. [DOI] [PubMed] [Google Scholar]
  36. Weller S. K., Temin H. M. Cell killing by avian leukosis viruses. J Virol. 1981 Sep;39(3):713–721. doi: 10.1128/jvi.39.3.713-721.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yu S. F., Baldwin D. N., Gwynn S. R., Yendapalli S., Linial M. L. Human foamy virus replication: a pathway distinct from that of retroviruses and hepadnaviruses. Science. 1996 Mar 15;271(5255):1579–1582. doi: 10.1126/science.271.5255.1579. [DOI] [PubMed] [Google Scholar]
  38. Yu S. F., Edelmann K., Strong R. K., Moebes A., Rethwilm A., Linial M. L. The carboxyl terminus of the human foamy virus Gag protein contains separable nucleic acid binding and nuclear transport domains. J Virol. 1996 Dec;70(12):8255–8262. doi: 10.1128/jvi.70.12.8255-8262.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Zack J. A., Arrigo S. J., Weitsman S. R., Go A. S., Haislip A., Chen I. S. HIV-1 entry into quiescent primary lymphocytes: molecular analysis reveals a labile, latent viral structure. Cell. 1990 Apr 20;61(2):213–222. doi: 10.1016/0092-8674(90)90802-l. [DOI] [PubMed] [Google Scholar]

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

RESOURCES