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Evidence that HIV-1 Rev directly promotes the nuclear export of unspliced RNA.

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Affiliations

  • 1. Institut für Molekularbiologie und Tumorforschung, Marburg, Germany.
      Authors
    • Fischer U 1
    (1 author)

The EMBO Journal, 01 Sep 1994, 13(17):4105-4112
https://doi.org/10.1002/j.1460-2075.1994.tb06728.x PMID: 8076606 PMCID: PMC395333

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Abstract 

The Rev trans-activator of human immunodeficiency virus type 1 (HIV-1) is a protein that regulates the simultaneous appearance in the cytoplasm of both spliced and unspliced forms of viral mRNAs from the same viral transcripts by way of recognition of a target sequence termed the Rev-responsive element (RRE). Whether Rev acts directly on RNA export or by inhibition of splicing, or both, is still a matter of debate. We have addressed this issue in Xenopus laevis oocytes by microinjecting RNA molecules containing RRE along with purified recombinant Rev protein into the oocyte nuclei. Adenovirus pre-mRNA containing an RRE in the intron was spliced equally well in the absence and presence of Rev protein. Only in the presence of Rev was non-spliced pre-mRNA exported from the nucleus; more surprisingly, the excised intron lariat (containing RRE) was also exported. Furthermore, an RRE-containing mRNA molecule that lacked intron sequences was also efficiently exported from the nucleus in a Rev-dependent manner. Therefore our results demonstrate that Rev can act directly at the level of nuclear export, independent of any inhibitory effect that it may exert on the splicing of pre-mRNA. Finally, our finding that the Rev mutant M10, shown previously to be inactive in human lymphoid cells, was also unable to export RRE-containing RNA molecules from oocyte nuclei suggests that one or more cellular factors, evolutionarily conserved between humans and Xenopus, interact with Rev in both cell systems to promote nuclear RNA export.

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Logo of embojLink to Publisher's site
EMBO J. 1994 Sep 1; 13(17): 4105–4112.
PMCID: PMC395333
PMID: 8076606

Evidence that HIV-1 Rev directly promotes the nuclear export of unspliced RNA.

U Fischer, S Meyer, M Teufel, C Heckel, R Lührmann, and G Rautmann
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Abstract

The Rev trans-activator of human immunodeficiency virus type 1 (HIV-1) is a protein that regulates the simultaneous appearance in the cytoplasm of both spliced and unspliced forms of viral mRNAs from the same viral transcripts by way of recognition of a target sequence termed the Rev-responsive element (RRE). Whether Rev acts directly on RNA export or by inhibition of splicing, or both, is still a matter of debate. We have addressed this issue in Xenopus laevis oocytes by microinjecting RNA molecules containing RRE along with purified recombinant Rev protein into the oocyte nuclei. Adenovirus pre-mRNA containing an RRE in the intron was spliced equally well in the absence and presence of Rev protein. Only in the presence of Rev was non-spliced pre-mRNA exported from the nucleus; more surprisingly, the excised intron lariat (containing RRE) was also exported. Furthermore, an RRE-containing mRNA molecule that lacked intron sequences was also efficiently exported from the nucleus in a Rev-dependent manner. Therefore our results demonstrate that Rev can act directly at the level of nuclear export, independent of any inhibitory effect that it may exert on the splicing of pre-mRNA. Finally, our finding that the Rev mutant M10, shown previously to be inactive in human lymphoid cells, was also unable to export RRE-containing RNA molecules from oocyte nuclei suggests that one or more cellular factors, evolutionarily conserved between humans and Xenopus, interact with Rev in both cell systems to promote nuclear RNA export.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Arrigo SJ, Chen IS. Rev is necessary for translation but not cytoplasmic accumulation of HIV-1 vif, vpr, and env/vpu 2 RNAs. Genes Dev. 1991 May;5(5):808–819. [Abstract] [Google Scholar]
  • Berger J, Aepinus C, Dobrovnik M, Fleckenstein B, Hauber J, Böhnlein E. Mutational analysis of functional domains in the HIV-1 Rev trans-regulatory protein. Virology. 1991 Aug;183(2):630–635. [Abstract] [Google Scholar]
  • Chang DD, Sharp PA. Regulation by HIV Rev depends upon recognition of splice sites. Cell. 1989 Dec 1;59(5):789–795. [Abstract] [Google Scholar]
  • Chang DD, Sharp PA. Messenger RNA transport and HIV rev regulation. Science. 1990 Aug 10;249(4969):614–615. [Abstract] [Google Scholar]
  • Cullen BR. Mechanism of action of regulatory proteins encoded by complex retroviruses. Microbiol Rev. 1992 Sep;56(3):375–394. [Europe PMC free article] [Abstract] [Google Scholar]
  • D'Agostino DM, Felber BK, Harrison JE, Pavlakis GN. The Rev protein of human immunodeficiency virus type 1 promotes polysomal association and translation of gag/pol and vpu/env mRNAs. Mol Cell Biol. 1992 Mar;12(3):1375–1386. [Europe PMC free article] [Abstract] [Google Scholar]
  • Daly TJ, Rennert P, Lynch P, Barry JK, Dundas M, Rusche JR, Doten RC, Auer M, Farrington GK. Perturbation of the carboxy terminus of HIV-1 Rev affects multimerization on the Rev responsive element. Biochemistry. 1993 Aug 31;32(34):8945–8954. [Abstract] [Google Scholar]
  • Dargemont C, Kühn LC. Export of mRNA from microinjected nuclei of Xenopus laevis oocytes. J Cell Biol. 1992 Jul;118(1):1–9. [Europe PMC free article] [Abstract] [Google Scholar]
  • Emerman M, Vazeux R, Peden K. The rev gene product of the human immunodeficiency virus affects envelope-specific RNA localization. Cell. 1989 Jun 30;57(7):1155–1165. [Abstract] [Google Scholar]
  • Feinberg MB, Jarrett RF, Aldovini A, Gallo RC, Wong-Staal F. HTLV-III expression and production involve complex regulation at the levels of splicing and translation of viral RNA. Cell. 1986 Sep 12;46(6):807–817. [Abstract] [Google Scholar]
  • Felber BK, Hadzopoulou-Cladaras M, Cladaras C, Copeland T, Pavlakis GN. rev protein of human immunodeficiency virus type 1 affects the stability and transport of the viral mRNA. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1495–1499. [Europe PMC free article] [Abstract] [Google Scholar]
  • Frendewey D, Keller W. Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences. Cell. 1985 Aug;42(1):355–367. [Abstract] [Google Scholar]
  • Hamm J, Mattaj IW. An abundant U6 snRNP found in germ cells and embryos of Xenopus laevis. EMBO J. 1989 Dec 20;8(13):4179–4187. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hamm J, Mattaj IW. Monomethylated cap structures facilitate RNA export from the nucleus. Cell. 1990 Oct 5;63(1):109–118. [Abstract] [Google Scholar]
  • Hamm J, Dathan NA, Mattaj IW. Functional analysis of mutant Xenopus U2 snRNAs. Cell. 1989 Oct 6;59(1):159–169. [Abstract] [Google Scholar]
  • Jarmolowski A, Boelens WC, Izaurralde E, Mattaj IW. Nuclear export of different classes of RNA is mediated by specific factors. J Cell Biol. 1994 Mar;124(5):627–635. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kjems J, Sharp PA. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kjems J, Brown M, Chang DD, Sharp PA. Structural analysis of the interaction between the human immunodeficiency virus Rev protein and the Rev response element. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):683–687. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kjems J, Frankel AD, Sharp PA. Specific regulation of mRNA splicing in vitro by a peptide from HIV-1 Rev. Cell. 1991 Oct 4;67(1):169–178. [Abstract] [Google Scholar]
  • Malim MH, Cullen BR. HIV-1 structural gene expression requires the binding of multiple Rev monomers to the viral RRE: implications for HIV-1 latency. Cell. 1991 Apr 19;65(2):241–248. [Abstract] [Google Scholar]
  • Malim MH, Cullen BR. Rev and the fate of pre-mRNA in the nucleus: implications for the regulation of RNA processing in eukaryotes. Mol Cell Biol. 1993 Oct;13(10):6180–6189. [Europe PMC free article] [Abstract] [Google Scholar]
  • Malim MH, Böhnlein S, Hauber J, Cullen BR. Functional dissection of the HIV-1 Rev trans-activator--derivation of a trans-dominant repressor of Rev function. Cell. 1989 Jul 14;58(1):205–214. [Abstract] [Google Scholar]
  • Malim MH, Hauber J, Le SY, Maizel JV, Cullen BR. The HIV-1 rev trans-activator acts through a structured target sequence to activate nuclear export of unspliced viral mRNA. Nature. 1989 Mar 16;338(6212):254–257. [Abstract] [Google Scholar]
  • Malim MH, McCarn DF, Tiley LS, Cullen BR. Mutational definition of the human immunodeficiency virus type 1 Rev activation domain. J Virol. 1991 Aug;65(8):4248–4254. [Europe PMC free article] [Abstract] [Google Scholar]
  • McDonald D, Hope TJ, Parslow TG. Posttranscriptional regulation by the human immunodeficiency virus type 1 Rev and human T-cell leukemia virus type I Rex proteins through a heterologous RNA binding site. J Virol. 1992 Dec;66(12):7232–7238. [Europe PMC free article] [Abstract] [Google Scholar]
  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. [Europe PMC free article] [Abstract] [Google Scholar]
  • Mermer B, Felber BK, Campbell M, Pavlakis GN. Identification of trans-dominant HIV-1 rev protein mutants by direct transfer of bacterially produced proteins into human cells. Nucleic Acids Res. 1990 Apr 25;18(8):2037–2044. [Europe PMC free article] [Abstract] [Google Scholar]
  • Muesing MA, Smith DH, Cabradilla CD, Benton CV, Lasky LA, Capon DJ. Nucleic acid structure and expression of the human AIDS/lymphadenopathy retrovirus. Nature. 1985 Feb 7;313(6002):450–458. [Abstract] [Google Scholar]
  • Mullis KB, Faloona FA. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. [Abstract] [Google Scholar]
  • Ruhl M, Himmelspach M, Bahr GM, Hammerschmid F, Jaksche H, Wolff B, Aschauer H, Farrington GK, Probst H, Bevec D, et al. Eukaryotic initiation factor 5A is a cellular target of the human immunodeficiency virus type 1 Rev activation domain mediating trans-activation. J Cell Biol. 1993 Dec;123(6 Pt 1):1309–1320. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schwartz S, Felber BK, Pavlakis GN. Distinct RNA sequences in the gag region of human immunodeficiency virus type 1 decrease RNA stability and inhibit expression in the absence of Rev protein. J Virol. 1992 Jan;66(1):150–159. [Europe PMC free article] [Abstract] [Google Scholar]
  • Terns MP, Dahlberg JE, Lund E. Multiple cis-acting signals for export of pre-U1 snRNA from the nucleus. Genes Dev. 1993 Oct;7(10):1898–1908. [Abstract] [Google Scholar]
  • Venkatesan S, Gerstberger SM, Park H, Holland SM, Nam Y. Human immunodeficiency virus type 1 Rev activation can be achieved without Rev-responsive element RNA if Rev is directed to the target as a Rev/MS2 fusion protein which tethers the MS2 operator RNA. J Virol. 1992 Dec;66(12):7469–7480. [Europe PMC free article] [Abstract] [Google Scholar]
  • Venkatesh LK, Chinnadurai G. Mutants in a conserved region near the carboxy-terminus of HIV-1 Rev identify functionally important residues and exhibit a dominant negative phenotype. Virology. 1990 Sep;178(1):327–330. [Abstract] [Google Scholar]
  • Zapp ML, Green MR. Sequence-specific RNA binding by the HIV-1 Rev protein. Nature. 1989 Dec 7;342(6250):714–716. [Abstract] [Google Scholar]
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