1887

Abstract

Antisense RNA, which has a sequence complementary to mRNA, may provide the basis for antiviral therapies of high selectivity. We have explored the inhibitory effect of six antisense RNAs upon the replication of human immunodeficiency virus (HIV) in cell culture. We chose regions of the HIV genome to test whether sequences required for splicing or for translation initiation were more susceptible to antisense RNA interference. Our results suggest that inhibitory antisense RNAs contain sequences complementary to the AUG initiation codon of the gene and have a comparatively low tendency to form intramolecular base pairs which would interfere with intermolecular duplex formation. Inhibition can be substantial (over 70%) but is transient. Transience does not result from mutation of the input virus. Inhibition was not a consequence of the induction of interferon by antisense RNA-mRNA duplex formation. Our results suggest that at least part of the inhibitory effect is at the posttranscriptional level.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-9-1965
1990-09-01
2021-10-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/9/JV0710091965.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-9-1965&mimeType=html&fmt=ahah

References

  1. Agrawal S., Ggodchild J., Civeira M. P., Thornton A. H., Sarin P. S., Zamecnik P. C. 1988; Oligodeoxynucleoside phosphoramidates and phosphorothioates as inhibitors of HIV. Proceedings of the National Academy of Sciences, U.S.A 85:7079–7083
    [Google Scholar]
  2. Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., Struhl K. 1987 Current Protocols in Molecular Biology New York: John Wiley;
    [Google Scholar]
  3. Bednarik D. P., Mosca J. D., Raj N. B., Pitha P. M. 1989; Inhibition of human immunodeficiency virus (HIV) replication by HIV-trans-activated alpha 2-interferon. Proceedings of the National Academy of Sciences, U.S.A 86:4958–4962
    [Google Scholar]
  4. Brown A. M. C., Scott M. R. D., Glover E. M. 1987; Retroviral Vectors. In DNA Cloning: A Practical Approach Ill: pp. 189–212 Glover D. M. Edited by Oxford: IRL Press;
    [Google Scholar]
  5. Chang L. J., Stoltzfus C. M. 1987; Inhibition of Rous sarcoma virus replication by antisense RNA. Journal of Virology 61:921–924
    [Google Scholar]
  6. Cotten M., Schaffner G., Birnstiel M. L. 1989; Ribozyme, antisense RNA, and antisense DNA inhibition of U7 small nuclear ribonucleoprotein-mediated histone pre-messenger RNA processing in vitro . Molecular and Cellular Biology 9:4479–4487
    [Google Scholar]
  7. Crowe S. M., McGrath M. S., Elbeik T., Kirihara J., Mills J. 1989; Comparative assessment of anti-retrovirals in human monocyte-macrophages and lymphoid cell lines acutely and chronically infected with the human immunodeficiency virus. Journal of Medical Virology 29:176–180
    [Google Scholar]
  8. Crowley T. E., Nellen W., Gomer R. H., Firtel R. A. 1985; Phenocopy of discoidin I-minus mutants by antisense transformation in Dictyostelium . Cell 43:633–641
    [Google Scholar]
  9. Cuozzo M., O’Connell K. M., Kaniewski W., Fang R. X., Chua N. H., Tumer N. E. 1988; Viral protection in transgenic tobacco plants expressing the cucumber mosaic virus coat protein or its antisense RNA. Bio/Technology 6:549
    [Google Scholar]
  10. Fisher A. G., Collalti E., Ratner L., Gallo R. C., Wong-Staal F. 1985; A molecular clone of HTLV-III with biological activity. Nature; London: 316262–265
    [Google Scholar]
  11. Foley G. E., Lazarus H., Farber S., Uzman B. G., Boone B. A., McCarthy R. E. 1965; Continuous culture of human lymphoblasts from peripheral blood of a child with acute leukemia. Cancer 18:522–529
    [Google Scholar]
  12. Frankel A. D., Pabo C. O. 1988; Cellular uptake of the tat protein from human immunodeficiency virus. Cell 55:1189–1193
    [Google Scholar]
  13. Freier S. M., Kierzek R., Jaeger J. A., Sugimoto N., Caruthers M. H., Neilson T., Turner D. H. 1986; Improved free-energy parameters for predictions of RNA duplex stability. Proceedings of the National Academy of Sciences, U.S.A 83:9373–9377
    [Google Scholar]
  14. Giebelhaus D. H., Eib D. W., Moon R. T. 1988; Antisense RNA inhibits expression of membrane skeleton protein 4.1 during embryonic development of Xenopus . Cel! 53:601–615
    [Google Scholar]
  15. Goodchild J., Agrawal S., Civeira M. P., Sarin P. S., Sun D., Zamecnik P. C. 1988; Inhibition of human immunodeficiency virus replication by antisense oligodeoxynucleotides. Proceedings of the National Academy of Sciences, U.S.A 85:5507–5551
    [Google Scholar]
  16. Gordon Y. J., Johnson B., Romanowski E., Araullocruz T. 1988; RNA complementary to herpes simplex virus type-1 ICP0 gene demonstrated in neurons of human trigeminal ganglia. Journal of Virology 62:1832–1835
    [Google Scholar]
  17. Hahn B. H., Shaw G. M., Arya S. K., Popovic M., Gallo R. C., Wong-Staal F. 1984; Molecular cloning and characterization of the HTLV-III virus associated with AIDS. Nature; London: 312166–169
    [Google Scholar]
  18. Hambor J. E., Hauer C. A., Shu H. -K., Groger R. K., Kaplan D. R. 1988; Use of an Epstein-Barr virus episomal replicon for anti-sense RNA-mediated gene inhibition in a human cytotoxic T-cell clone. Proceedings of the National Academy of Sciences, U.S.A 85:4010–4014
    [Google Scholar]
  19. Hemenway C., Fang R. X., Kaniewski W. K., Chua N. H., Tumer N. E. 1988; Analysis of the mechanism of protection in transgenic plants expressing the potato virus-X coat protein or its antisense RNA. EMBO Journal 7:1273–1280
    [Google Scholar]
  20. Izant J. G., Weintraub H. 1984; Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis. Cell 36:1007–1015
    [Google Scholar]
  21. Jainchill J., Aaronson S. A., Todaro G. J. 1969; Murine sarcoma and leukemia viruses: assay using clonal lines of contact- inhibited mouse cells. Journal of Virology 4:549–553
    [Google Scholar]
  22. Jennings P. A., Molloy P. L. 1987; Inhibition of SV40 replicon function by engineered antisense RNA transcribed by RNA polymerase III. EMBO Journal 6:3043–3047
    [Google Scholar]
  23. Kerr S. M., Stark G. R., Kerr I. M. 1988; Excess antisense RNA from infectious recombinant SV40 fails to inhibit expression of a transfected, interferon-inducible gene. European Journal of Biochemistry 175:65–73
    [Google Scholar]
  24. Khochbin S., Lawrence J. -J. 1989; An antisense RNA involved in p53 mRNA maturation in murine erythroleukemia cells induced to differentiate. EMBO Journal 8:4107–4114
    [Google Scholar]
  25. Kim S. K., Wold B. J. 1985; Stable reduction of thymidine kinase activity in cells expressing high levels of anti-sense RNA. Cell 42:129–138
    [Google Scholar]
  26. Kimelman D., Kirschner M. W. 1989; An antisense mRNA directs the covalent modification of the transcript encoding fibroblast growth factor in Xenopus oocytes. Cell 59:687–696
    [Google Scholar]
  27. Koka P., Yunis J., Passarelli A. L., Dubey D. P., Faller D. V., Yunis E. J. 1988; Increased expression of CD4 molecules on Jurkat cells mediated by human immunodeficiency virus Tat protein. Journal of Virology 62:4353–4357
    [Google Scholar]
  28. McGarry T. J., Lindquist S. 1986; Inhibition of heat shock protein synthesis by heat-inducible antisense RNA. Proceedings of the National Academy of Sciences, U.S.A 83:399–403
    [Google Scholar]
  29. McMichael A. 1987 Leukocyte Typing 3: Oxford: Oxford University Press;
    [Google Scholar]
  30. Mann R., Mulligan R. C., Baltimore D. 1983; Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell 33:153–159
    [Google Scholar]
  31. Matsukura M., Shinozuka K., Zon G., Mitsuya H., Reitz M., Cohen J. S., Broder S. 1987; Phosphorothioate analogs of oligodeoxynucleotides: inhibitors of replication and cytopathic effects of human immunodeficiency virus. Proceedings of the National Academy of Sciences, U.S.A 84:7706–7710
    [Google Scholar]
  32. Miller A. D., Buttimore C. 1986; Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Molecular and Cellular Biology 6:2895–2902
    [Google Scholar]
  33. Munroe S. H. 1988; Antisense RNA inhibits splicing of pre-mRNA in vitro . EMBO Journal 7:2523–2532
    [Google Scholar]
  34. Nakamaye K., Eckstein F. 1986; Inhibition of restriction endonuclease Ncil cleavage by phosphorothioate groups and its application to oligonucleotide-directed mutagenesis. Nucleic Acids Research 14:9679–9698
    [Google Scholar]
  35. Ratner L., Haseltine W., Patarca R., Livak K. J., Starcich B., Josephs S. F., Doran E. R., Rafalski J. A., Whitehorn E. A., Baumeister K., Ivanoff L., Petteway S. R., Pearson M. L., Lautenberger J. A., Papas T. S., Ghrayeb J., Chang N. T., Gallo R. C., Wong-Staal F. 1985; Complete nucleotide sequence of the AIDS virus. Nature; London: 313277–284
    [Google Scholar]
  36. Rodermel S. R., Abbott M. S., Bogorad L. 1988; Nuclear-organelle interactions: nuclear antisense gene inhibits ribulosebisphosphate carboxylase enzyme levels in transformed tobacco plants. Cell 55:673–681
    [Google Scholar]
  37. Rogers J. C. 1988; RNA complementary to alpha-amylase mRNA in barley. Plant Molecular Biology 11:125–138
    [Google Scholar]
  38. Salser W. 1978; Globin mRNA sequences: analysis of base pairing and evolutionary implications. Cold Spring Harbor Symposia on Quantitative Biology 42:985–1002
    [Google Scholar]
  39. Sandri-Goldin R. M., Sekulovich R. E., Leary K. 1987; The alpha protein, ICP0, does not appear to play a major role in the regulation of herpes simplex virus gene expression during infection in tissue culture. Nucleic Acids Research 15:905–919
    [Google Scholar]
  40. Sarin P., Agrawal S., Civeira M. P., Goodchild J., Ikeuchi T., Zamecnik P. C. 1988; Inhibition of AIDS virus by oligodeoxy- nucleoside methyl phosphonates. Proceedings of the National Academy of Sciences, U.S.A 85:7448–7451
    [Google Scholar]
  41. Shibahara S., Mukai S., Morisawa H., Nakashima H., Kobaya-shi S., Yamamoto N. 1989; Inhibition of human immunodeficiency virus (HIV-1) replication by synthetic oligo-RNA derivatives. Nucleic Acids Research 17:239–252
    [Google Scholar]
  42. Simons R. W., Kleckner N. 1988; Biological regulation by antisense RNA in prokaryotes. Annual Review of Genetics 22:567–601
    [Google Scholar]
  43. Smith C. J. S., Watson C. F., Ray J., Bird C. R., Morris P. C., Schuch W., Grierson D. 1988; Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature; London: 334:724
    [Google Scholar]
  44. Taylor J. W., Ott J., Eckstein F. 1985; The rapid generation of oligonucleotide-directed mutations at high frequency using phos- phorothioate-modified DNA. Nucleic Acids Research 13:8764–8785
    [Google Scholar]
  45. Volk R., Koster M., Poting A., Hartmann L., Knochel W. 1989; An antisense transcript from the XenopuslaevisbFGF gene coding for an evolutionarily conserved 24kd protein. EMBO Journal 8:2983–2988
    [Google Scholar]
  46. Von Ruden T., Gilboa E. 1989; Inhibition of human T-cell leukaemia virus type 1 replication in primary human T cells that express antisense RNA. Journal of Virology 63:677–682
    [Google Scholar]
  47. Walder J. 1988; Antisense DNA and RNA: progress and prospects. Genes and Development 2:502–504
    [Google Scholar]
  48. Weiss A., Wiskocil R. L., Stobo J. D. 1984; The role of T3 surface molecules in the activation of human T cells: a two stimulus requirement for IL-2 production reflects events occurring at a pretranslational level. Journal of Immunology 133:123–128
    [Google Scholar]
  49. Yamada O., Hattori N., Kurimura T., Kita M., Kishida T. 1988; Inhibition of growth of HIV by human natural interferon in vitro . AIDS Research and Human Retroviruses 4:287–294
    [Google Scholar]
  50. Zaia J. A., Rossi J. J., Murakawa G. J., Spallone P. A., Stephens D. A., Kaplan B. E., Eritia R., Wallace R. B., Cantin E. M. 1988; Inhibition of human immunodeficiency virus by using an oligonucleoside methyl-phosphonate targeted to the tat-3 gene. Journal of Virology 62:3914–3917
    [Google Scholar]
  51. Zamecnik P. C., Goodchild J., Tguchi Y., Sarin P. 1986; Inhibition of replication and expression of human T-cell lymphotro- pic virus type III in cultured cells by exogenous synthetic oligonucleotides complementary to viral RNA. Proceedings of the National Academy of Sciences, U.S.A 83:4143–4146
    [Google Scholar]
  52. Zuker M. 1989; Computer prediction of RNA structure. Methods in Enzymology 180:262–288
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-71-9-1965
Loading
/content/journal/jgv/10.1099/0022-1317-71-9-1965
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error