An important aspect of the pathophysiology of human immunodeficiency virus type-1 (HIV-1) infection is the ability of the virus to replicate in non-dividing cells1,2,3. HIV-1 matrix (MA), the amino-terminal domain of the Pr55 gag polyprotein (Pr55), bears a nuclear localization signal that promotes localization of the viral preintegration complex to the nucleus of non-dividing cells following virus entry3,4,5. However, late during infection, MA, as part of Pr55, directs unspliced viral RNA to the plasma membrane6, the site of virus assembly. How MA can mediate these two opposing targeting functions is not understood. Here we demonstrate that MA has a previously undescribed nuclear export activity. Although MA lacks the canonical leucine-rich nuclear export signal, nuclear export is mediated through the conserved Crmlp pathway and functions in both mammalian cells and yeast. A mutation that disrupts the MA nuclear export signal (MA-M4) mislocalizes Pr55 and genomic viral RNA to the nucleus, thereby severely impairing viral replication. Furthermore, we show that MA-M4 can act in a dominant-negative fashion to mislocalize genomic viral RNA even in the presence of wild-type MA. We conclude that the MA nuclear export signal is required to counteract the MA nuclear localization signal, thus ensuring the cytoplasmic availability of the components required for virion assembly.
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Lewis,P., Hensel,M. & Emerman,M. Human immunodeficiency virus infection of cells arrested in the cell cycle. EMBO J. 11, 3053–3058 (1992).
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. 174, 1477–1482 (1991).
Bukrinksy,M. I. et al. Active nuclear import of human immunodeficiency virus type 1 preintegration complexes. Proc. Natl Acad. Sci. USA 89, 6580–6584 (1992).
Bukrinsky,M. I. et al. A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells. Nature 365, 666–669 (1993).
Bukrinsky,M. I. et al. Association of integrase, matrix, and reverse transcriptase antigens of human immunodeficiency virus type 1 with viral nucleic acids following acute infection. Proc. Natl Acad. Sci. USA 90, 6125–6129 (1993).
Krausslich,H. G. & Welker,R. Intracellular transport of retroviral capsid components. Curr. Top. Microbiol. Immunol. 214, 25–63 (1996).
Fisher,A. G. et al. The trans-activator gene of HTLV-III is essential for virus replication. Nature 320, 367–371 (1986).
Freed,E. O. HIV-1 gag proteins: diverse functions in the virus life cycle. Virology 251, 1–15 (1998).
Clever,J. L. & Parslow,T. G. Mutant human immunodeficiency virus type 1 genomes with defects in RNA dimerization or encapsidation. J. Virol. 71, 3407–3414 (1997).
Paillart,J. C. et al. A dual role of the putative RNA dimerization initiation site of human immunodeficiency virus type 1 in genomic RNA packaging and proviral DNA synthesis. J. Virol. 70, 8348–8354 (1996).
Matthews,S. et al. Structural similarity between the p17 matrix protein of HIV-1 and interferon-gamma. Nature 370, 666–668 (1994).
Massiah,M. A. et al. Three-dimensional structure of the human immunodeficiency virus type 1 matrix protein. J. Mol. Biol. 244, 198–223 (1994).
Wolff,B., Sanglier,J. J. & Wang,Y. Leptomycin B is an inhibitor of nuclear export: inhibition of nucleo-cytoplasmic translocation of the human immunodeficiency virus type 1 (HIV-1) Rev protein and Rev-dependent mRNA. Chem. Biol. 4, 139–147 (1997).
Nishi,K. et al. Leptomycin B targets a regulatory cascade of crm1, a fission yeast nuclear protein, involved in control of higher order chromosome structure and gene expression. J. Biol. Chem. 269, 6320–6324 (1994).
Fornerod,M., Ohno,M., Yoshida,M. & Mattaj,I. W. CRM1 is an export receptor for leucine-rich nuclear export signals. Cells 90, 1051–1060 (1997).
Fukuda,M. et al. CRM1 is responsible for intracellular transport mediated by the nuclear export signal. Nature 390, 308–311 (1997).
Ossareh-Nazari,B., Bacheleric,F. & Dargemont,C. Evidence for a role of CRM1 in signal-mediated nuclear protein export. Science 278, 141–144 (1997).
Stade,K., Ford,C. S., Guthrie,C. & Weis,K. Exportin 1 (Crm1p) is an essential nuclear export factor. Cell 90, 1041–1050 (1997).
Berkowitz,R., Fisher,J. & Goff,S. P. RNA Packaging. Curr. Top. Microb. Immunol. 214, 177–218 (1996).
von Schwedler,U., Kornbluth,R. S. & Trono,D. The nuclear localization signal of the matrix protein of human immunodeficiency virus type 1 allows the establishment of infection in macrophages and quiescent T lymphocytes. Proc. Natl Acad. Sci. USA 91, 6992–6996 (1994).
Fouchier,R. A., Meyer,B. E., Simon,J. H., Fischer,U. & Malim,M. H. HIV-1 infection of non-dividing cells: evidence that the amino-terminal basic region of the viral matrix protein is important for Gag processing but not for post-entry nuclear import. EMBO J. 16, 4531–4539 (1997).
Freed,E. O., Englund,G. & Martin,M. A. Role of the basic domain of human immunodeficiency virus type 1 matrix in macrophage infection. J. Virol. 69, 3949–3954 (1995).
Chomczynski,P. A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques 15, 532–534 (1993).
Sakuragi,J. I. & Panganiban,A. T. Human immunodeficiency virus type 1 RNA outside the primary encapsidation and dimer linkage region affects RNA dimer stability in vivo. J. Virol. 71, 3250–3254 (1997).
Kislauskis,E. H., Zhu,X. & Singer,R. H. Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype. J. Cell Biol. 127, 441–451 (1994).
Neville,M., Stutz,F., Lee,L., Davis,L. I. & Rosbash,M. The importin-beta family member crm1p bridges the interaction between Rev and the nuclear pore complex during nuclear export. Curr. Biol. 7, 767–775 (1997).
We thank J. Mondor, N. Bakker and L. Ross for manuscript preparation. We also thank M. Yoshida for generous donation of Leptomycin B, and M. Rosbash for yeast strains. We thank J. Kan, J. Teodoro, M. Sharkey, J.-M. Jacque, B. Brichacek, S. Swingler and M. Zapp for scientific discussions. This work was supported in part by grants from the NIH to M.R.G. and M.S., and A.B. is supported in part by a Fogarty International Research Collaboration Award. M.R.G. is an investigator of the Howard Hughes Medical Institute.
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Dupont, S., Sharova, N., DéHoratius, C. et al. A novel nuclear export activity in HIV-1 matrix protein required for viral replication. Nature 402, 681–685 (1999) doi:10.1038/45272
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