Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1995 Jan;69(1):348–356. doi: 10.1128/jvi.69.1.348-356.1995

Development of physical forms of unintegrated retroviral DNA in mouse spinal cord tissue during ts1-induced spongiform encephalomyelopathy: elevated levels of a novel single-stranded form in paralyzed mice.

P F Szurek 1, B R Brooks 1
PMCID: PMC188582  PMID: 7983729

Abstract

ts1 is a murine leukemia virus that causes rapidly evolving hindlimb paralysis in susceptible strains of mice. Following perinatal infection, three physical forms of unintegrated viral DNA were detected in the spinal cord by Southern blot hybridization. Linear and supercoiled closed-circle viral double-stranded DNAs were detected in both the central nervous system and non-central nervous system tissues. An elevated level of a novel minus-sense single-stranded form of viral DNA, which had a very high mobility in agarose gels, was correlated with the onset of symptoms of paralysis. As the severity of paralysis progressed, the level of this single-stranded form increased rapidly, with the highest level in the spinal cords of moribund mice. Since the virulence of a number of cytopathic retroviruses has been associated with the presence of increased amounts of unintegrated viral DNA in the tissues of the infected hosts, this novel form of highly mobile unintegrated single-stranded DNA may have a role in the neuropathogenesis of ts1.

Full Text

The Full Text of this article is available as a PDF (427.4 KB).

Selected References

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

  1. Arkhipova I. R., Mazo A. M., Cherkasova V. A., Gorelova T. V., Schuppe N. G., Llyin Y. V. The steps of reverse transcription of Drosophila mobile dispersed genetic elements and U3-R-U5 structure of their LTRs. Cell. 1986 Feb 28;44(4):555–563. doi: 10.1016/0092-8674(86)90265-5. [DOI] [PubMed] [Google Scholar]
  2. Ball J., McCarter J. A., Sunderland S. M. Evidence for helper independent murine sarcoma virus. I. Segregation of replication-defective and transformation-defective viruses. Virology. 1973 Nov;56(1):268–284. doi: 10.1016/0042-6822(73)90305-x. [DOI] [PubMed] [Google Scholar]
  3. Baszler T. V., Zachary J. F. Murine retroviral neurovirulence correlates with an enhanced ability ofvirus to infect selectively, replicate in, and activate resident microglial cells. Am J Pathol. 1991 Mar;138(3):655–671. [PMC free article] [PubMed] [Google Scholar]
  4. Buckler C. E., Hoggan M. D., Chan H. W., Sears J. F., Khan A. S., Moore J. L., Hartley J. W., Rowe W. P., Martin M. A. Cloning and characterization of an envelope-specific probe from xenotropic murine leukemia proviral DNA. J Virol. 1982 Jan;41(1):228–236. doi: 10.1128/jvi.41.1.228-236.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buller R. S., Wehrly K., Portis J. L., Chesebro B. Host genes conferring resistance to a central nervous system disease induced by a polytropic recombinant Friend murine retrovirus. J Virol. 1990 Feb;64(2):493–498. doi: 10.1128/jvi.64.2.493-498.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chan H. W., Bryan T., Moore J. L., Staal S. P., Rowe W. P., Martin M. A. Identification of ecotropic proviral sequences in inbred mouse strains with a cloned subgenomic DNA fragment. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5779–5783. doi: 10.1073/pnas.77.10.5779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Donahue P. R., Quackenbush S. L., Gallo M. V., deNoronha C. M., Overbaugh J., Hoover E. A., Mullins J. I. Viral genetic determinants of T-cell killing and immunodeficiency disease induction by the feline leukemia virus FeLV-FAIDS. J Virol. 1991 Aug;65(8):4461–4469. doi: 10.1128/jvi.65.8.4461-4469.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  9. Freed E. O., Risser R. The role of envelope glycoprotein processing in murine leukemia virus infection. J Virol. 1987 Sep;61(9):2852–2856. doi: 10.1128/jvi.61.9.2852-2856.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Henderson L. E., Sowder R., Copeland T. D., Smythers G., Oroszlan S. Quantitative separation of murine leukemia virus proteins by reversed-phase high-pressure liquid chromatography reveals newly described gag and env cleavage products. J Virol. 1984 Nov;52(2):492–500. doi: 10.1128/jvi.52.2.492-500.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Herr W., Schwartz D., Gilbert W. Isolation and mapping of cDNA hybridization probes specific for ecotropic and nonecotropic murine leukemia proviruses. Virology. 1983 Feb;125(1):139–154. doi: 10.1016/0042-6822(83)90070-3. [DOI] [PubMed] [Google Scholar]
  12. Holter W., Rabson A. B., Corsico C. D., Howard B. H. Sequence-specific toxicity of transfected retroviral DNA. Exp Cell Res. 1991 Mar;193(1):54–58. doi: 10.1016/0014-4827(91)90537-5. [DOI] [PubMed] [Google Scholar]
  13. Horiuchi K., Zinder N. D. Site-specific cleavage of single-stranded DNA by a Hemophilus restriction endonuclease. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2555–2558. doi: 10.1073/pnas.72.7.2555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jaenisch R. Retroviruses and embryogenesis: microinjection of Moloney leukemia virus into midgestation mouse embryos. Cell. 1980 Jan;19(1):181–188. doi: 10.1016/0092-8674(80)90399-2. [DOI] [PubMed] [Google Scholar]
  15. Jolicoeur P. Neuronal loss in a lower motor neuron disease induced by a murine retrovirus. Can J Neurol Sci. 1991 Aug;18(3 Suppl):411–413. doi: 10.1017/s031716710003256x. [DOI] [PubMed] [Google Scholar]
  16. Jones R. W., Jones M. J. Simplified filter paper sandwich blot provides rapid, background-free northern blots. Biotechniques. 1992 May;12(5):684–688. [PubMed] [Google Scholar]
  17. Kay D. G., Gravel C., Pothier F., Laperrière A., Robitaille Y., Jolicoeur P. Neurological disease induced in transgenic mice expressing the env gene of the Cas-Br-E murine retrovirus. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4538–4542. doi: 10.1073/pnas.90.10.4538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kim J. W., Closs E. I., Albritton L. M., Cunningham J. M. Transport of cationic amino acids by the mouse ecotropic retrovirus receptor. Nature. 1991 Aug 22;352(6337):725–728. doi: 10.1038/352725a0. [DOI] [PubMed] [Google Scholar]
  19. Lazo P. A., Prasad V., Tsichlis P. N. Splice acceptor site for the env message of Moloney murine leukemia virus. J Virol. 1987 Jun;61(6):2038–2041. doi: 10.1128/jvi.61.6.2038-2041.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Majors J. E., Varmus H. E. Nucleotide sequencing of an apparent proviral copy of env mRNA defines determinants of expression of the mouse mammary tumor virus env gene. J Virol. 1983 Sep;47(3):495–504. doi: 10.1128/jvi.47.3.495-504.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maniatis T., Jeffrey A., van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. doi: 10.1021/bi00688a010. [DOI] [PubMed] [Google Scholar]
  22. McCarter J. A., Ball J. K., Frei J. V. Lower limb paralysis induced in mice by a temperature-sensitive mutant of Moloney leukemia virus. J Natl Cancer Inst. 1977 Jul;59(1):179–183. doi: 10.1093/jnci/59.1.179. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Nagra R. M., Wong P. K., Wiley C. A. Expression of major histocompatibility complex antigens and serum neutralizing antibody in murine retroviral encephalitis. J Neuropathol Exp Neurol. 1993 Mar;52(2):163–173. doi: 10.1097/00005072-199303000-00009. [DOI] [PubMed] [Google Scholar]
  25. O'Neill R. R., Khan A. S., Hoggan M. D., Hartley J. W., Martin M. A., Repaske R. Specific hybridization probes demonstrate fewer xenotropic than mink cell focus-forming murine leukemia virus env-related sequences in DNAs from inbred laboratory mice. J Virol. 1986 May;58(2):359–366. doi: 10.1128/jvi.58.2.359-366.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Oldstone M. B., Jensen F., Elder J., Dixon F. J., Lampert P. W. Pathogenesis of the slow disease of the central nervous system associated with wild mouse virus. III. Role of input virus and MCF recombinants in disease. Virology. 1983 Jul 15;128(1):154–165. doi: 10.1016/0042-6822(83)90326-4. [DOI] [PubMed] [Google Scholar]
  27. Orita M., Iwahana H., Kanazawa H., Hayashi K., Sekiya T. Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2766–2770. doi: 10.1073/pnas.86.8.2766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pang S., Koyanagi Y., Miles S., Wiley C., Vinters H. V., Chen I. S. High levels of unintegrated HIV-1 DNA in brain tissue of AIDS dementia patients. Nature. 1990 Jan 4;343(6253):85–89. doi: 10.1038/343085a0. [DOI] [PubMed] [Google Scholar]
  29. Reinhart T. A., Ghosh A. K., Hoover E. A., Mullins J. I. Distinct superinfection interference properties yet similar receptor utilization by cytopathic and noncytopathic feline leukemia viruses. J Virol. 1993 Sep;67(9):5153–5162. doi: 10.1128/jvi.67.9.5153-5162.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roe T., Reynolds T. C., Yu G., Brown P. O. Integration of murine leukemia virus DNA depends on mitosis. EMBO J. 1993 May;12(5):2099–2108. doi: 10.1002/j.1460-2075.1993.tb05858.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rogers S. G., Weiss B. Exonuclease III of Escherichia coli K-12, an AP endonuclease. Methods Enzymol. 1980;65(1):201–211. doi: 10.1016/s0076-6879(80)65028-9. [DOI] [PubMed] [Google Scholar]
  32. Schwartzberg P. L., Roth M. J., Tanese N., Goff S. P. Analysis of a temperature-sensitive mutation affecting the integration protein of Moloney murine leukemia virus. Virology. 1993 Feb;192(2):673–678. doi: 10.1006/viro.1993.1086. [DOI] [PubMed] [Google Scholar]
  33. Shikova E., Lin Y. C., Saha K., Brooks B. R., Wong P. K. Correlation of specific virus-astrocyte interactions and cytopathic effects induced by ts1, a neurovirulent mutant of Moloney murine leukemia virus. J Virol. 1993 Mar;67(3):1137–1147. doi: 10.1128/jvi.67.3.1137-1147.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Stansly P. G. Non-oncogenic infectious agents associated with experimental tumors. Prog Exp Tumor Res. 1965;7:224–258. doi: 10.1159/000391382. [DOI] [PubMed] [Google Scholar]
  37. Steffen D., Bird S., Rowe W. P., Weinberg R. A. Identification of DNA fragments carrying ecotropic proviruses of AKR mice. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4554–4558. doi: 10.1073/pnas.76.9.4554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Stoica G., Floyd E., Illanes O., Wong P. K. Temporal lymphoreticular changes caused by ts1, a paralytogenic mutant of Moloney murine leukemia virus TB. Lab Invest. 1992 Apr;66(4):427–436. [PubMed] [Google Scholar]
  39. Stoica G., Illanes O., Tasca S. I., Wong P. K. Temporal central and peripheral nervous system changes induced by a paralytogenic mutant of Moloney murine leukemia virus TB. Lab Invest. 1993 Dec;69(6):724–735. [PubMed] [Google Scholar]
  40. Szurek P. F., Floyd E., Yuen P. H., Wong P. K. Site-directed mutagenesis of the codon for Ile-25 in gPr80env alters the neurovirulence of ts1, a mutant of Moloney murine leukemia virus TB. J Virol. 1990 Nov;64(11):5241–5249. doi: 10.1128/jvi.64.11.5241-5249.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Szurek P. F., Yuen P. H., Ball J. K., Wong P. K. A Val-25-to-Ile substitution in the envelope precursor polyprotein, gPr80env, is responsible for the temperature sensitivity, inefficient processing of gPr80env, and neurovirulence of ts1, a mutant of Moloney murine leukemia virus TB. J Virol. 1990 Feb;64(2):467–475. doi: 10.1128/jvi.64.2.467-475.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Szurek P. F., Yuen P. H., Jerzy R., Wong P. K. Identification of point mutations in the envelope gene of Moloney murine leukemia virus TB temperature-sensitive paralytogenic mutant ts1: molecular determinants for neurovirulence. J Virol. 1988 Jan;62(1):357–360. doi: 10.1128/jvi.62.1.357-360.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Taddeo B., Federico M., Titti F., Rossi G. B., Verani P. Homologous superinfection of both producer and nonproducer HIV-infected cells is blocked at a late retrotranscription step. Virology. 1993 Jun;194(2):441–452. doi: 10.1006/viro.1993.1283. [DOI] [PubMed] [Google Scholar]
  44. Taketo M., Schroeder A. C., Mobraaten L. E., Gunning K. B., Hanten G., Fox R. R., Roderick T. H., Stewart C. L., Lilly F., Hansen C. T. FVB/N: an inbred mouse strain preferable for transgenic analyses. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2065–2069. doi: 10.1073/pnas.88.6.2065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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]
  46. Temin H. M. Retrovirus variation and reverse transcription: abnormal strand transfers result in retrovirus genetic variation. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):6900–6903. doi: 10.1073/pnas.90.15.6900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. 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]
  48. Varela-Echavarría A., Garvey N., Preston B. D., Dougherty J. P. Comparison of Moloney murine leukemia virus mutation rate with the fidelity of its reverse transcriptase in vitro. J Biol Chem. 1992 Dec 5;267(34):24681–24688. [PubMed] [Google Scholar]
  49. 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]
  50. Vaux D. L. Toward an understanding of the molecular mechanisms of physiological cell death. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):786–789. doi: 10.1073/pnas.90.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Weller S. K., Joy A. E., Temin H. M. Correlation between cell killing and massive second-round superinfection by members of some subgroups of avian leukosis virus. J Virol. 1980 Jan;33(1):494–506. doi: 10.1128/jvi.33.1.494-506.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wiley C. A., Gardner M. The pathogenesis of murine retroviral infection of the central nervous system. Brain Pathol. 1993 Apr;3(2):123–128. doi: 10.1111/j.1750-3639.1993.tb00736.x. [DOI] [PubMed] [Google Scholar]
  53. Wong P. K., Knupp C., Yuen P. H., Soong M. M., Zachary J. F., Tompkins W. A. ts1, a Paralytogenic mutant of Moloney murine leukemia virus TB, has an enhanced ability to replicate in the central nervous system and primary nerve cell culture. J Virol. 1985 Sep;55(3):760–767. doi: 10.1128/jvi.55.3.760-767.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wong P. K., Russ L. J., McCarter J. A. Rapid, selective procedure for isolation of spontaneous temperature-sensitive mutants of Moloney leukemia virus. Virology. 1973 Feb;51(2):424–431. doi: 10.1016/0042-6822(73)90441-8. [DOI] [PubMed] [Google Scholar]
  55. Wong P. K., Soong M. M., MacLeod R., Gallick G. E., Yuen P. H. A group of temperature-sensitive mutants of Moloney leukemia virus which is defective in cleavage of env precursor polypeptide in infected cells also induces hind-limb paralysis in newborn CFW/D mice. Virology. 1983 Mar;125(2):513–518. doi: 10.1016/0042-6822(83)90225-8. [DOI] [PubMed] [Google Scholar]
  56. Yang W. K., Kiggans J. O., Yang D. M., Ou C. Y., Tennant R. W., Brown A., Bassin R. H. Synthesis and circularization of N- and B-tropic retroviral DNA Fv-1 permissive and restrictive mouse cells. Proc Natl Acad Sci U S A. 1980 May;77(5):2994–2998. doi: 10.1073/pnas.77.5.2994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Yoshimura F. K., Weinberg R. A. Restriction endonuclease cleavage of linear and closed circular murine leukemia viral DNAs: discovery of a smaller circular form. Cell. 1979 Feb;16(2):323–332. doi: 10.1016/0092-8674(79)90009-6. [DOI] [PubMed] [Google Scholar]
  58. Yuen P. H., Malehorn D., Knupp C., Wong P. K. A 1.6-kilobase-pair fragment in the genome of the ts1 mutant of Moloney murine leukemia virus TB that is associated with temperature sensitivity, nonprocessing of Pr80env, and paralytogenesis. J Virol. 1985 May;54(2):364–373. doi: 10.1128/jvi.54.2.364-373.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Yuen P. H., Malehorn D., Nau C., Soong M. M., Wong P. K. Molecular cloning of two paralytogenic, temperature-sensitive mutants, ts1 and ts7, and the parental wild-type Moloney murine leukemia virus. J Virol. 1985 Apr;54(1):178–185. doi: 10.1128/jvi.54.1.178-185.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Yuen P. H., Szurek P. F. The reduced virulence of the thymotropic Moloney murine leukemia virus derivative MoMuLV-TB is mapped to 11 mutations within the U3 region of the long terminal repeat. J Virol. 1989 Feb;63(2):471–480. doi: 10.1128/jvi.63.2.471-480.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Zachary J. F., Knupp C. J., Wong P. K. Noninflammatory spongiform polioencephalomyelopathy caused by a neurotropic temperature-sensitive mutant of Moloney murine leukemia virus TB. Am J Pathol. 1986 Sep;124(3):457–468. [PMC free article] [PubMed] [Google Scholar]
  62. 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]
  63. Zhu J., Cunningham J. M. Minus-strand DNA is present within murine type C ecotropic retroviruses prior to infection. J Virol. 1993 Apr;67(4):2385–2388. doi: 10.1128/jvi.67.4.2385-2388.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES