Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1996 Aug;70(8):5511–5518. doi: 10.1128/jvi.70.8.5511-5518.1996

Human T-cell leukemia virus type 2 Rex inhibits pre-mRNA splicing in vitro at an early stage of spliceosome formation.

A Bakker 1, X Li 1, C T Ruland 1, D W Stephens 1, A C Black 1, J D Rosenblatt 1
PMCID: PMC190509  PMID: 8764063

Abstract

The Rex protein is an essential regulator of RNA expression in human T-cell leukemia virus types 1 and 2 (HTLV-1 and HTLV-2) that promotes the accumulation of full-length and partially spliced viral transcripts in the cytoplasm. Rex-mediated regulation correlates with specific binding to a cognate RNA recognition element which overlaps the 5' splice site in the viral long terminal repeat. It has been unclear whether Rex directly affects splicing or only nuclear-to-cytoplasmic transport of viral mRNA. We demonstrate that HTLV-2 Rex is a potent inhibitor of splicing in vitro at an early step in spliceosome assembly. Inhibition requires phosphorylation of Rex and the ability of Rex to bind to the Rex response element. Direct inhibition of early spliceosome assembly by Rex may account for differential accumulation of unspliced transcripts and represents a novel mechanism of retroviral gene regulation.

Full Text

The Full Text of this article is available as a PDF (561.2 KB).

Selected References

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

  1. Arrigo S., Beemon K. Regulation of Rous sarcoma virus RNA splicing and stability. Mol Cell Biol. 1988 Nov;8(11):4858–4867. doi: 10.1128/mcb.8.11.4858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Black A. C., Chen I. S., Arrigo S., Ruland C. T., Allogiamento T., Chin E., Rosenblatt J. D. Regulation of HTLV-II gene expression by Rex involves positive and negative cis-acting elements in the 5'long terminal repeat. Virology. 1991 Apr;181(2):433–444. doi: 10.1016/0042-6822(91)90875-c. [DOI] [PubMed] [Google Scholar]
  3. Black A. C., Ruland C. T., Luo J., Bakker A., Fraser J. K., Rosenblatt J. D. Binding of nuclear proteins to HTLV-II cis-acting repressive sequence (CRS) RNA correlates with CRS function. Virology. 1994 Apr;200(1):29–41. doi: 10.1006/viro.1994.1159. [DOI] [PubMed] [Google Scholar]
  4. Black A. C., Ruland C. T., Yip M. T., Luo J., Tran B., Kalsi A., Quan E., Aboud M., Chen I. S., Rosenblatt J. D. Human T-cell leukemia virus type II Rex binding and activity require an intact splice donor site and a specific RNA secondary structure. J Virol. 1991 Dec;65(12):6645–6653. doi: 10.1128/jvi.65.12.6645-6653.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang D. D., Sharp P. A. Regulation by HIV Rev depends upon recognition of splice sites. Cell. 1989 Dec 1;59(5):789–795. doi: 10.1016/0092-8674(89)90602-8. [DOI] [PubMed] [Google Scholar]
  6. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fischer U., Meyer S., Teufel M., Heckel C., Lührmann R., Rautmann G. Evidence that HIV-1 Rev directly promotes the nuclear export of unspliced RNA. EMBO J. 1994 Sep 1;13(17):4105–4112. doi: 10.1002/j.1460-2075.1994.tb06728.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gontarek R. R., McNally M. T., Beemon K. Mutation of an RSV intronic element abolishes both U11/U12 snRNP binding and negative regulation of splicing. Genes Dev. 1993 Oct;7(10):1926–1936. doi: 10.1101/gad.7.10.1926. [DOI] [PubMed] [Google Scholar]
  9. Green P. L., Xie Y. M., Chen I. S. The Rex proteins of human T-cell leukemia virus type II differ by serine phosphorylation. J Virol. 1991 Jan;65(1):546–550. doi: 10.1128/jvi.65.1.546-550.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Green P. L., Yip M. T., Xie Y., Chen I. S. Phosphorylation regulates RNA binding by the human T-cell leukemia virus Rex protein. J Virol. 1992 Jul;66(7):4325–4330. doi: 10.1128/jvi.66.7.4325-4330.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hidaka M., Inoue J., Yoshida M., Seiki M. Post-transcriptional regulator (rex) of HTLV-1 initiates expression of viral structural proteins but suppresses expression of regulatory proteins. EMBO J. 1988 Feb;7(2):519–523. doi: 10.1002/j.1460-2075.1988.tb02840.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Inoue J., Itoh M., Akizawa T., Toyoshima H., Yoshida M. HTLV-1 Rex protein accumulates unspliced RNA in the nucleus as well as in cytoplasm. Oncogene. 1991 Oct;6(10):1753–1757. [PubMed] [Google Scholar]
  13. Katz R. A., Skalka A. M. Control of retroviral RNA splicing through maintenance of suboptimal processing signals. Mol Cell Biol. 1990 Feb;10(2):696–704. doi: 10.1128/mcb.10.2.696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kim J. H., Kaufman P. A., Hanly S. M., Rimsky L. T., Greene W. C. Rex transregulation of human T-cell leukemia virus type II gene expression. J Virol. 1991 Jan;65(1):405–414. doi: 10.1128/jvi.65.1.405-414.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kjems J., Sharp P. A. The basic domain of Rev from human immunodeficiency virus type 1 specifically blocks the entry of U4/U6.U5 small nuclear ribonucleoprotein in spliceosome assembly. J Virol. 1993 Aug;67(8):4769–4776. doi: 10.1128/jvi.67.8.4769-4776.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Konarska M. M., Sharp P. A. Interactions between small nuclear ribonucleoprotein particles in formation of spliceosomes. Cell. 1987 Jun 19;49(6):763–774. doi: 10.1016/0092-8674(87)90614-3. [DOI] [PubMed] [Google Scholar]
  17. Lewis N., Williams J., Rekosh D., Hammarskjöld M. L. Identification of a cis-acting element in human immunodeficiency virus type 2 (HIV-2) that is responsive to the HIV-1 rev and human T-cell leukemia virus types I and II rex proteins. J Virol. 1990 Apr;64(4):1690–1697. doi: 10.1128/jvi.64.4.1690-1697.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Malim M. H., Hauber J., Fenrick R., Cullen B. R. Immunodeficiency virus rev trans-activator modulates the expression of the viral regulatory genes. Nature. 1988 Sep 8;335(6186):181–183. doi: 10.1038/335181a0. [DOI] [PubMed] [Google Scholar]
  19. Nagashima K., Yoshida M., Seiki M. A single species of pX mRNA of human T-cell leukemia virus type I encodes trans-activator p40x and two other phosphoproteins. J Virol. 1986 Nov;60(2):394–399. doi: 10.1128/jvi.60.2.394-399.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nosaka T., Siomi H., Adachi Y., Ishibashi M., Kubota S., Maki M., Hatanaka M. Nucleolar targeting signal of human T-cell leukemia virus type I rex-encoded protein is essential for cytoplasmic accumulation of unspliced viral mRNA. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9798–9802. doi: 10.1073/pnas.86.24.9798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rimsky L., Hauber J., Dukovich M., Malim M. H., Langlois A., Cullen B. R., Greene W. C. Functional replacement of the HIV-1 rev protein by the HTLV-1 rex protein. Nature. 1988 Oct 20;335(6192):738–740. doi: 10.1038/335738a0. [DOI] [PubMed] [Google Scholar]
  22. Rosenblatt J. D., Cann A. J., Slamon D. J., Smalberg I. S., Shah N. P., Fujii J., Wachsman W., Chen I. S. HTLV-II transactivation is regulated by the overlapping tax/rex nonstructural genes. Science. 1988 May 13;240(4854):916–919. doi: 10.1126/science.2834826. [DOI] [PubMed] [Google Scholar]
  23. Seed B., Sheen J. Y. A simple phase-extraction assay for chloramphenicol acyltransferase activity. Gene. 1988 Jul 30;67(2):271–277. doi: 10.1016/0378-1119(88)90403-9. [DOI] [PubMed] [Google Scholar]
  24. Sharp P. A. Split genes and RNA splicing. Cell. 1994 Jun 17;77(6):805–815. doi: 10.1016/0092-8674(94)90130-9. [DOI] [PubMed] [Google Scholar]
  25. Yip M. T., Dynan W. S., Green P. L., Black A. C., Arrigo S. J., Torbati A., Heaphy S., Ruland C., Rosenblatt J. D., Chen I. S. Human T-cell leukemia virus (HTLV) type II Rex protein binds specifically to RNA sequences of the HTLV long terminal repeat but poorly to the human immunodeficiency virus type 1 Rev-responsive element. J Virol. 1991 May;65(5):2261–2272. doi: 10.1128/jvi.65.5.2261-2272.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zhang L., Stoltzfus C. M. A suboptimal src 3' splice site is necessary for efficient replication of Rous sarcoma virus. Virology. 1995 Feb 1;206(2):1099–1107. doi: 10.1006/viro.1995.1033. [DOI] [PubMed] [Google Scholar]

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

RESOURCES