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. 1993 May;12(5):2099–2108. doi: 10.1002/j.1460-2075.1993.tb05858.x

Integration of murine leukemia virus DNA depends on mitosis.

T Roe 1, T C Reynolds 1, G Yu 1, P O Brown 1
PMCID: PMC413431  PMID: 8491198

Abstract

In synchronized rat or mouse cells infected with Moloney murine leukemia virus (MLV), integration of viral DNA and production of viral proteins occur only after the cells traverse mitosis. Integration is blocked when cells are prevented from progressing through mitosis. Viral nucleoprotein complexes isolated from arrested cells contain full-length viral DNA and can integrate this viral DNA in vitro, showing that the block to integration in arrested cells is not due to a lack of mature integration machinery. When infected cells traverse mitosis, there is a sharp increase in nuclear accumulation of viral DNA. The dependence of integration on mitosis may therefore be due to a requirement for mitosis and nuclear envelope breakdown for entry of the viral integration complex into the nucleus.

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Selected References

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

  1. Bader J. P. A change in growth potential of cells after conversion by Rous sarcoma virus. J Cell Physiol. 1967 Dec;70(3):301–308. doi: 10.1002/jcp.1040700310. [DOI] [PubMed] [Google Scholar]
  2. Bader J. P. Metabolic requirements for infection by Rous sarcoma virus. IV. Virus reproduction and cellular transformation without cellular division. Virology. 1972 May;48(2):494–501. doi: 10.1016/0042-6822(72)90060-8. [DOI] [PubMed] [Google Scholar]
  3. Batterson W., Furlong D., Roizman B. Molecular genetics of herpes simplex virus. VIII. further characterization of a temperature-sensitive mutant defective in release of viral DNA and in other stages of the viral reproductive cycle. J Virol. 1983 Jan;45(1):397–407. doi: 10.1128/jvi.45.1.397-407.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blow J. J., Laskey R. A. A role for the nuclear envelope in controlling DNA replication within the cell cycle. Nature. 1988 Apr 7;332(6164):546–548. doi: 10.1038/332546a0. [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. Brown P. O., Bowerman B., Varmus H. E., Bishop J. M. Correct integration of retroviral DNA in vitro. Cell. 1987 May 8;49(3):347–356. doi: 10.1016/0092-8674(87)90287-x. [DOI] [PubMed] [Google Scholar]
  7. Brown P. O., Bowerman B., Varmus H. E., Bishop J. M. Retroviral integration: structure of the initial covalent product and its precursor, and a role for the viral IN protein. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2525–2529. doi: 10.1073/pnas.86.8.2525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bukrinsky M. I., Sharova N., Dempsey M. P., Stanwick T. L., Bukrinskaya A. G., Haggerty S., Stevenson M. Active nuclear import of human immunodeficiency virus type 1 preintegration complexes. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6580–6584. doi: 10.1073/pnas.89.14.6580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Byrnes J. J. Structural and functional properties of DNA polymerase delta from rabbit bone marrow. Mol Cell Biochem. 1984 Apr;62(1):13–24. doi: 10.1007/BF00230073. [DOI] [PubMed] [Google Scholar]
  10. Chida K., Vogt P. K. Nuclear translocation of viral Jun but not of cellular Jun is cell cycle dependent. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4290–4294. doi: 10.1073/pnas.89.10.4290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Clever J., Yamada M., Kasamatsu H. Import of simian virus 40 virions through nuclear pore complexes. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7333–7337. doi: 10.1073/pnas.88.16.7333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dales S., Chardonnet Y. Early events in the interaction of adenoviruses with HeLa cells. IV. Association with microtubules and the nuclear pore complex during vectorial movement of the inoculum. Virology. 1973 Dec;56(2):465–483. doi: 10.1016/0042-6822(73)90050-0. [DOI] [PubMed] [Google Scholar]
  13. De Brabander M. J., Van de Veire R. M., Aerts F. E., Borgers M., Janssen P. A. The effects of methyl (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl) carbamate, (R 17934; NSC 238159), a new synthetic antitumoral drug interfering with microtubules, on mammalian cells cultured in vitro. Cancer Res. 1976 Mar;36(3):905–916. [PubMed] [Google Scholar]
  14. 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]
  15. Fritsch E., Temin H. M. Formation and structure of infectious DNA of spleen necrosis virus. J Virol. 1977 Jan;21(1):119–130. doi: 10.1128/jvi.21.1.119-130.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gallant P., Nigg E. A. Cyclin B2 undergoes cell cycle-dependent nuclear translocation and, when expressed as a non-destructible mutant, causes mitotic arrest in HeLa cells. J Cell Biol. 1992 Apr;117(1):213–224. doi: 10.1083/jcb.117.1.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Guntaka R. V., Richards O. C., Shank P. R., Kung H. J., Davidson N. Covalently closed circular DNA of avian sarcoma virus: purification from nuclei of infected quail tumor cells and measurement by electron microscopy and gel electrophoresis. J Mol Biol. 1976 Sep 15;106(2):337–357. doi: 10.1016/0022-2836(76)90090-5. [DOI] [PubMed] [Google Scholar]
  18. Harel J., Rassart E., Jolicoeur P. Cell cycle dependence of synthesis of unintegrated viral DNA in mouse cells newly infected with murine leukemia virus. Virology. 1981 Apr 15;110(1):202–207. doi: 10.1016/0042-6822(81)90022-2. [DOI] [PubMed] [Google Scholar]
  19. Hartmann C., Golde A., Villaudy J., Park I., Vernet G., Krsmanovic V. Study of pp60v-src protein kinase activity in synchronized chicken embryo fibroblasts infected with Rous sarcoma virus. Exp Cell Res. 1984 Jan;150(1):242–249. doi: 10.1016/0014-4827(84)90719-5. [DOI] [PubMed] [Google Scholar]
  20. Hennessy K. M., Clark C. D., Botstein D. Subcellular localization of yeast CDC46 varies with the cell cycle. Genes Dev. 1990 Dec;4(12B):2252–2263. doi: 10.1101/gad.4.12b.2252. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Hoebeke J., Van Nijen G., De Brabander M. Interaction of oncodazole (R 17934), a new antitumoral drug, with rat brain tubulin. Biochem Biophys Res Commun. 1976 Mar 22;69(2):319–324. doi: 10.1016/0006-291x(76)90524-6. [DOI] [PubMed] [Google Scholar]
  23. Hsu T. W., Taylor J. M. Effect of aphidicolin on avian sarcoma virus replication. J Virol. 1982 Nov;44(2):493–498. doi: 10.1128/jvi.44.2.493-498.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hummeler K., Tomassini N., Sokol F. Morphological aspects of the uptake of simian virus 40 by permissive cells. J Virol. 1970 Jul;6(1):87–93. doi: 10.1128/jvi.6.1.87-93.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Humphries E. H., Glover C., Reichmann M. E. Rous sarcoma virus infection of synchronized cells establishes provirus integration during S-phase DNA synthesis prior to cellular division. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2601–2605. doi: 10.1073/pnas.78.4.2601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Humphries E. H., Temin H. M. Cell cycle-dependent activation of rous sarcoma virus-infected stationary chicken cells: avian leukosis virus group-specific antigens and ribonucleic acid. J Virol. 1972 Jul;10(1):82–87. doi: 10.1128/jvi.10.1.82-87.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Humphries E. H., Temin H. M. Requirement for cell division for initiation of transcription of Rous sarcoma virus RNA. J Virol. 1974 Sep;14(3):531–546. doi: 10.1128/jvi.14.3.531-546.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Johnson G. D., Nogueira Araujo G. M. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981;43(3):349–350. doi: 10.1016/0022-1759(81)90183-6. [DOI] [PubMed] [Google Scholar]
  29. Jolicoeur P., Baltimore D. Effect of Fv-1 gene product on proviral DNA formation and integration in cells infected with murine leukemia viruses. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2236–2240. doi: 10.1073/pnas.73.7.2236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Jolicoeur P., Rassart E. Effect of Fv-1 gene product on synthesis of linear and supercoiled viral DNA in cells infected with murine leukemia virus. J Virol. 1980 Jan;33(1):183–195. doi: 10.1128/jvi.33.1.183-195.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Jolicoeur P. The Fv-1 gene of the mouse and its control of murine leukemia virus replication. Curr Top Microbiol Immunol. 1979;86:67–122. doi: 10.1007/978-3-642-67341-2_3. [DOI] [PubMed] [Google Scholar]
  32. Leno G. H., Downes C. S., Laskey R. A. The nuclear membrane prevents replication of human G2 nuclei but not G1 nuclei in Xenopus egg extract. Cell. 1992 Apr 3;69(1):151–158. doi: 10.1016/0092-8674(92)90126-w. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Lobel L. I., Patel M., King W., Nguyen-Huu M. C., Goff S. P. Construction and recovery of viable retroviral genomes carrying a bacterial suppressor transfer RNA gene. Science. 1985 Apr 19;228(4697):329–332. doi: 10.1126/science.2984770. [DOI] [PubMed] [Google Scholar]
  35. Martin K., Helenius A. Transport of incoming influenza virus nucleocapsids into the nucleus. J Virol. 1991 Jan;65(1):232–244. doi: 10.1128/jvi.65.1.232-244.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Martinez J., Georgoff I., Martinez J., Levine A. J. Cellular localization and cell cycle regulation by a temperature-sensitive p53 protein. Genes Dev. 1991 Feb;5(2):151–159. doi: 10.1101/gad.5.2.151. [DOI] [PubMed] [Google Scholar]
  37. McMorrow I., Souter W. E., Plopper G., Burke B. Identification of a Golgi-associated protein that undergoes mitosis dependent phosphorylation and relocation. J Cell Biol. 1990 May;110(5):1513–1523. doi: 10.1083/jcb.110.5.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Miller D. G., Adam M. A., Miller A. D. Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol Cell Biol. 1990 Aug;10(8):4239–4242. doi: 10.1128/mcb.10.8.4239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Mineo C., Murakami Y., Ishimi Y., Hanaoka F., Yamada M. Isolation and analysis of a mammalian temperature-sensitive mutant defective in G2 functions. Exp Cell Res. 1986 Nov;167(1):53–62. doi: 10.1016/0014-4827(86)90203-x. [DOI] [PubMed] [Google Scholar]
  40. Miyamoto K., Morgan C. Structure and development of viruses as observed in the electron microscope. XI. Entry and uncoating of herpes simplex virus. J Virol. 1971 Dec;8(6):910–918. doi: 10.1128/jvi.8.6.910-918.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Moll T., Tebb G., Surana U., Robitsch H., Nasmyth K. The role of phosphorylation and the CDC28 protein kinase in cell cycle-regulated nuclear import of the S. cerevisiae transcription factor SWI5. Cell. 1991 Aug 23;66(4):743–758. doi: 10.1016/0092-8674(91)90118-i. [DOI] [PubMed] [Google Scholar]
  42. Nigg E. A., Baeuerle P. A., Lührmann R. Nuclear import-export: in search of signals and mechanisms. Cell. 1991 Jul 12;66(1):15–22. doi: 10.1016/0092-8674(91)90135-l. [DOI] [PubMed] [Google Scholar]
  43. Peters R. Fluorescence microphotolysis to measure nucleocytoplasmic transport and intracellular mobility. Biochim Biophys Acta. 1986 Dec 22;864(3-4):305–359. doi: 10.1016/0304-4157(86)90003-1. [DOI] [PubMed] [Google Scholar]
  44. Pines J., Hunter T. Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport. J Cell Biol. 1991 Oct;115(1):1–17. doi: 10.1083/jcb.115.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Pryciak P. M., Sil A., Varmus H. E. Retroviral integration into minichromosomes in vitro. EMBO J. 1992 Jan;11(1):291–303. doi: 10.1002/j.1460-2075.1992.tb05052.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Pryciak P. M., Varmus H. E. Fv-1 restriction and its effects on murine leukemia virus integration in vivo and in vitro. J Virol. 1992 Oct;66(10):5959–5966. doi: 10.1128/jvi.66.10.5959-5966.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. RUBIN H., TEMIN H. M. A radiological study of cell-virus interaction in the Rous sarcoma. Virology. 1959 Jan;7(1):75–91. doi: 10.1016/0042-6822(59)90178-3. [DOI] [PubMed] [Google Scholar]
  48. Shank P. R., Varmus H. E. Virus-specific DNA in the cytoplasm of avian sarcoma virus-infected cells is a precursor to covalently closed circular viral DNA in the nucleus. J Virol. 1978 Jan;25(1):104–104. doi: 10.1128/jvi.25.1.104-104.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Springett G. M., Moen R. C., Anderson S., Blaese R. M., Anderson W. F. Infection efficiency of T lymphocytes with amphotropic retroviral vectors is cell cycle dependent. J Virol. 1989 Sep;63(9):3865–3869. doi: 10.1128/jvi.63.9.3865-3869.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Stevenson M., Stanwick T. L., Dempsey M. P., Lamonica C. A. HIV-1 replication is controlled at the level of T cell activation and proviral integration. EMBO J. 1990 May;9(5):1551–1560. doi: 10.1002/j.1460-2075.1990.tb08274.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Summers M. D. Electron microscopic observations on granulosis virus entry, uncoating and replication processes during infection of the midgut cells of Trichoplusia ni. J Ultrastruct Res. 1971 Jun;35(5):606–625. doi: 10.1016/s0022-5320(71)80014-x. [DOI] [PubMed] [Google Scholar]
  52. Th'ng J. P., Wright P. S., Hamaguchi J., Lee M. G., Norbury C. J., Nurse P., Bradbury E. M. The FT210 cell line is a mouse G2 phase mutant with a temperature-sensitive CDC2 gene product. Cell. 1990 Oct 19;63(2):313–324. doi: 10.1016/0092-8674(90)90164-a. [DOI] [PubMed] [Google Scholar]
  53. Tognon M., Furlong D., Conley A. J., Roizman B. Molecular genetics of herpes simplex virus. V. Characterization of a mutant defective in ability to form plaques at low temperatures and in a viral fraction which prevents accumulation of coreless capsids at nuclear pores late in infection. J Virol. 1981 Dec;40(3):870–880. doi: 10.1128/jvi.40.3.870-880.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Trask B., Pinkel D. Fluorescence in situ hybridization with DNA probes. Methods Cell Biol. 1990;33:383–400. doi: 10.1016/s0091-679x(08)60542-7. [DOI] [PubMed] [Google Scholar]
  55. Varmus H. E., Padgett T., Heasley S., Simon G., Bishop J. M. Cellular functions are required for the synthesis and integration of avian sarcoma virus-specific DNA. Cell. 1977 Jun;11(2):307–319. doi: 10.1016/0092-8674(77)90047-2. [DOI] [PubMed] [Google Scholar]
  56. Weinberg J. B., Matthews T. J., Cullen B. R., Malim M. H. Productive human immunodeficiency virus type 1 (HIV-1) infection of nonproliferating human monocytes. J Exp Med. 1991 Dec 1;174(6):1477–1482. doi: 10.1084/jem.174.6.1477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. 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]

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