Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1995 Jun;69(6):3675–3682. doi: 10.1128/jvi.69.6.3675-3682.1995

Nascent human immunodeficiency virus type 1 reverse transcription occurs within an enveloped particle.

H Zhang 1, Y Zhang 1, T Spicer 1, D Henrard 1, B J Poiesz 1
PMCID: PMC189083  PMID: 7745716

Abstract

Although a small amount of viral DNA has been shown to be enclosed within human immunodeficiency virus type 1 (HIV-1) virions, the majority of full-length viral DNA is formed after this virus infects target cells. Hence, we undertook investigations to identify the physical characteristics of the HIV-1 replication unit during the early events of infection. In these studies, nascent viral DNA synthesis was found to occur between 15 and 30 min after purified, DNase-treated HIV-1 virions were added to HUT 78 cells. At 1 h postinfection, a large amount of strong-stop viral DNA and some first-strand viral DNA had been synthesized. Several lines of evidence, including purification, nuclease digestion, and immunoprecipitation, indicated that these nascent viral DNAs were located within particles containing components such as reverse transcriptase and p24gag and gp120env proteins and having physical characteristics similar to those of intact virions.

Full Text

The Full Text of this article is available as a PDF (891.6 KB).

Selected References

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

  1. Abbott M. A., Poiesz B. J., Byrne B. C., Kwok S., Sninsky J. J., Ehrlich G. D. Enzymatic gene amplification: qualitative and quantitative methods for detecting proviral DNA amplified in vitro. J Infect Dis. 1988 Dec;158(6):1158–1169. doi: 10.1093/infdis/158.6.1158. [DOI] [PubMed] [Google Scholar]
  2. Adachi A., Gendelman H. E., Koenig S., Folks T., Willey R., Rabson A., Martin M. A. Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone. J Virol. 1986 Aug;59(2):284–291. doi: 10.1128/jvi.59.2.284-291.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Amin R. M., Dean M. T., Zaumetzer L. E., Poiesz B. J. Virucidal efficacy of various lens cleaning and disinfecting solutions on HIV-I contaminated contact lenses. AIDS Res Hum Retroviruses. 1991 Apr;7(4):403–408. doi: 10.1089/aid.1991.7.403. [DOI] [PubMed] [Google Scholar]
  4. Bagasra O., Farzadegan H., Seshamma T., Oakes J. W., Saah A., Pomerantz R. J. Detection of HIV-1 proviral DNA in sperm from HIV-1-infected men. AIDS. 1994 Dec;8(12):1669–1674. doi: 10.1097/00002030-199412000-00005. [DOI] [PubMed] [Google Scholar]
  5. Bowerman B., Brown P. O., Bishop J. M., Varmus H. E. A nucleoprotein complex mediates the integration of retroviral DNA. Genes Dev. 1989 Apr;3(4):469–478. doi: 10.1101/gad.3.4.469. [DOI] [PubMed] [Google Scholar]
  6. Bukrinskaya A. G., Sharova N. K. Unusual features of protein interaction in human immunodeficiency virus (HIV) virions. Arch Virol. 1990;110(3-4):287–293. doi: 10.1007/BF01311297. [DOI] [PubMed] [Google Scholar]
  7. Bukrinsky M. I., Sharova N., McDonald T. L., Pushkarskaya T., Tarpley W. G., Stevenson M. Association of integrase, matrix, and reverse transcriptase antigens of human immunodeficiency virus type 1 with viral nucleic acids following acute infection. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6125–6129. doi: 10.1073/pnas.90.13.6125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dyster L. M., Abbott L., Bryz-Gornia V., Poiesz B. J., Papsidero L. D. Microplate-based DNA hybridization assays for detection of human retroviral gene sequences. J Clin Microbiol. 1994 Feb;32(2):547–550. doi: 10.1128/jcm.32.2.547-550.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farnet C. M., Haseltine W. A. Determination of viral proteins present in the human immunodeficiency virus type 1 preintegration complex. J Virol. 1991 Apr;65(4):1910–1915. doi: 10.1128/jvi.65.4.1910-1915.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Farnet C. M., Haseltine W. A. Integration of human immunodeficiency virus type 1 DNA in vitro. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4164–4168. doi: 10.1073/pnas.87.11.4164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Folks T. M., Powell D., Lightfoote M., Koenig S., Fauci A. S., Benn S., Rabson A., Daugherty D., Gendelman H. E., Hoggan M. D. Biological and biochemical characterization of a cloned Leu-3- cell surviving infection with the acquired immune deficiency syndrome retrovirus. J Exp Med. 1986 Jul 1;164(1):280–290. doi: 10.1084/jem.164.1.280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Gazdar A. F., Carney D. N., Bunn P. A., Russell E. K., Jaffe E. S., Schechter G. P., Guccion J. G. Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas. Blood. 1980 Mar;55(3):409–417. [PubMed] [Google Scholar]
  14. Gelderblom H. R., Hausmann E. H., Ozel M., Pauli G., Koch M. A. Fine structure of human immunodeficiency virus (HIV) and immunolocalization of structural proteins. Virology. 1987 Jan;156(1):171–176. doi: 10.1016/0042-6822(87)90449-1. [DOI] [PubMed] [Google Scholar]
  15. Grewe C., Beck A., Gelderblom H. R. HIV: early virus-cell interactions. J Acquir Immune Defic Syndr. 1990;3(10):965–974. [PubMed] [Google Scholar]
  16. Henrard D. R., Mehaffey W. F., Allain J. P. A sensitive viral capture assay for detection of plasma viremia in HIV-infected individuals. AIDS Res Hum Retroviruses. 1992 Jan;8(1):47–52. doi: 10.1089/aid.1992.8.47. [DOI] [PubMed] [Google Scholar]
  17. Kaplan A. H., Krogstad P., Kempf D. J., Norbeck D. W., Swanstrom R. Human immunodeficiency virus type 1 virions composed of unprocessed Gag and Gag-Pol precursors are capable of reverse transcribing viral genomic RNA. Antimicrob Agents Chemother. 1994 Dec;38(12):2929–2933. doi: 10.1128/aac.38.12.2929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Karageorgos L., Li P., Burrell C. Characterization of HIV replication complexes early after cell-to-cell infection. AIDS Res Hum Retroviruses. 1993 Sep;9(9):817–823. doi: 10.1089/aid.1993.9.817. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Kwok S., Kellogg D., Ehrlich G., Poiesz B., Bhagavati S., Sninsky J. J. Characterization of a sequence of human T cell leukemia virus type I from a patient with chronic progressive myelopathy. J Infect Dis. 1988 Dec;158(6):1193–1197. doi: 10.1093/infdis/158.6.1193. [DOI] [PubMed] [Google Scholar]
  21. Kwok S., Mack D. H., Mullis K. B., Poiesz B., Ehrlich G., Blair D., Friedman-Kien A., Sninsky J. J. Identification of human immunodeficiency virus sequences by using in vitro enzymatic amplification and oligomer cleavage detection. J Virol. 1987 May;61(5):1690–1694. doi: 10.1128/jvi.61.5.1690-1694.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Layne S. P., Merges M. J., Dembo M., Spouge J. L., Conley S. R., Moore J. P., Raina J. L., Renz H., Gelderblom H. R., Nara P. L. Factors underlying spontaneous inactivation and susceptibility to neutralization of human immunodeficiency virus. Virology. 1992 Aug;189(2):695–714. doi: 10.1016/0042-6822(92)90593-e. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Maddon P. J., Dalgleish A. G., McDougal J. S., Clapham P. R., Weiss R. A., Axel R. The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain. Cell. 1986 Nov 7;47(3):333–348. doi: 10.1016/0092-8674(86)90590-8. [DOI] [PubMed] [Google Scholar]
  25. McEwen C. R. Tables for estimating sedimentation through linear concentration gradients of sucrose solution. Anal Biochem. 1967 Jul;20(1):114–149. doi: 10.1016/0003-2697(67)90271-0. [DOI] [PubMed] [Google Scholar]
  26. McKeating J. A., McKnight A., Moore J. P. Differential loss of envelope glycoprotein gp120 from virions of human immunodeficiency virus type 1 isolates: effects on infectivity and neutralization. J Virol. 1991 Feb;65(2):852–860. doi: 10.1128/jvi.65.2.852-860.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Oh S. K., Cruikshank W. W., Raina J., Blanchard G. C., Adler W. H., Walker J., Kornfeld H. Identification of HIV-1 envelope glycoprotein in the serum of AIDS and ARC patients. J Acquir Immune Defic Syndr. 1992;5(3):251–256. [PubMed] [Google Scholar]
  28. Pauza C. D., Price T. M. Human immunodeficiency virus infection of T cells and monocytes proceeds via receptor-mediated endocytosis. J Cell Biol. 1988 Sep;107(3):959–968. doi: 10.1083/jcb.107.3.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Peliska J. A., Balasubramanian S., Giedroc D. P., Benkovic S. J. Recombinant HIV-1 nucleocapsid protein accelerates HIV-1 reverse transcriptase catalyzed DNA strand transfer reactions and modulates RNase H activity. Biochemistry. 1994 Nov 22;33(46):13817–13823. doi: 10.1021/bi00250a036. [DOI] [PubMed] [Google Scholar]
  30. Poiesz B. J., Ruscetti F. W., Gazdar A. F., Bunn P. A., Minna J. D., Gallo R. C. Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7415–7419. doi: 10.1073/pnas.77.12.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Preston B. D., Poiesz B. J., Loeb L. A. Fidelity of HIV-1 reverse transcriptase. Science. 1988 Nov 25;242(4882):1168–1171. doi: 10.1126/science.2460924. [DOI] [PubMed] [Google Scholar]
  32. Ross E. K., Fuerst T. R., Orenstein J. M., O'Neill T., Martin M. A., Venkatesan S. Maturation of human immunodeficiency virus particles assembled from the gag precursor protein requires in situ processing by gag-pol protease. AIDS Res Hum Retroviruses. 1991 May;7(5):475–483. doi: 10.1089/aid.1991.7.475. [DOI] [PubMed] [Google Scholar]
  33. Sadaie M. R., Kalyanaraman V. S., Mukopadhayaya R., Tschachler E., Gallo R. C., Wong-Staal F. Biological characterization of noninfectious HIV-1 particles lacking the envelope protein. Virology. 1992 Apr;187(2):604–611. doi: 10.1016/0042-6822(92)90462-x. [DOI] [PubMed] [Google Scholar]
  34. Scofield V. L., Rao B., Broder S., Kennedy C., Wallace M., Graham B., Poiesz B. J. HIV interaction with sperm. AIDS. 1994 Dec;8(12):1733–1736. [PubMed] [Google Scholar]
  35. Stein B. S., Gowda S. D., Lifson J. D., Penhallow R. C., Bensch K. G., Engleman E. G. pH-independent HIV entry into CD4-positive T cells via virus envelope fusion to the plasma membrane. Cell. 1987 Jun 5;49(5):659–668. doi: 10.1016/0092-8674(87)90542-3. [DOI] [PubMed] [Google Scholar]
  36. Taswell C. Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis. J Immunol. 1981 Apr;126(4):1614–1619. [PubMed] [Google Scholar]
  37. 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]
  38. Varmus H. E., Heasley S., Kung H. J., Oppermann H., Smith V. C., Bishop J. M., Shank P. R. Kinetics of synthesis, structure and purification of avian sarcoma virus-specific DNA made in the cytoplasm of acutely infected cells. J Mol Biol. 1978 Mar 25;120(1):55–82. doi: 10.1016/0022-2836(78)90295-4. [DOI] [PubMed] [Google Scholar]
  39. Wells K. H., Byrne B. C., Poiesz B. J. Detection, prevention, and treatment of retroviral infections. Semin Oncol. 1990 Jun;17(3):295–320. [PubMed] [Google Scholar]
  40. Wells K. H., Latino J., Gavalchin J., Poiesz B. J. Inactivation of human immunodeficiency virus type 1 by ozone in vitro. Blood. 1991 Oct 1;78(7):1882–1890. [PubMed] [Google Scholar]
  41. 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]
  42. Zack J. A., Haislip A. M., Krogstad P., Chen I. S. Incompletely reverse-transcribed human immunodeficiency virus type 1 genomes in quiescent cells can function as intermediates in the retroviral life cycle. J Virol. 1992 Mar;66(3):1717–1725. doi: 10.1128/jvi.66.3.1717-1725.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zhang H., Bagasra O., Niikura M., Poiesz B. J., Pomerantz R. J. Intravirion reverse transcripts in the peripheral blood plasma on human immunodeficiency virus type 1-infected individuals. J Virol. 1994 Nov;68(11):7591–7597. doi: 10.1128/jvi.68.11.7591-7597.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zhang H., Zhang Y., Spicer T. P., Abbott L. Z., Abbott M., Poiesz B. J. Reverse transcription takes place within extracellular HIV-1 virions: potential biological significance. AIDS Res Hum Retroviruses. 1993 Dec;9(12):1287–1296. doi: 10.1089/aid.1993.9.1287. [DOI] [PubMed] [Google Scholar]

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

RESOURCES